From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!NMSU.EDU!hroychow From: hroychow@NMSU.EDU (Hiranya Roychowdhury) Newsgroups: bionet.molbio.proteins Subject: Re: Tissue homogenizer for Eppies? Date: 2 Jun 1997 14:22:15 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <199706022122.PAA55078@hector.NMSU.Edu> NNTP-Posting-Host: net.bio.net At 08:48 AM 6/2/97 +0000, Keld Sorensen wrote: >I think Kontes sell these with a little motorized pestle. > >Keld. > > We had gone ahead and bought one of those 'motoized' things. It is useless when you are grinding tissues frozen in liq N2. Dr. Hiranya Sankar Roychowdhury Plant Genetic Engineering Lab. New Mexico State University Las Cruces, NM 88003 Ph. (505) 646-5785 hroychow@nmsu.edu From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!ihnp4.ucsd.edu!swrinde!howland.erols.net!newshub2.home.com!newshub1.home.com!news.home.com!nick.arc.nasa.gov!purdue!mozo.cc.purdue.edu!news From: Ali McBride Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: Mon, 02 Jun 1997 14:01:52 -0500 Organization: BMS Lines: 25 Message-ID: <339318A0.270E@bilbo.bio.purdue.edu> References: <338EFCA7.41C6@risotto.mit.edu> Reply-To: amcbride@bilbo.bio.purdue.edu NNTP-Posting-Host: bms06.vet.purdue.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01Gold (Win95; U) Xref: biosci bionet.cellbiol:7418 bionet.immunology:11892 bionet.molbio.methds-reagnts:58205 bionet.molbio.proteins:10889 David Aldridge wrote: > > In article <338EFCA7.41C6@risotto.mit.edu>, Thomas Cameron > wrote: > > > Company kits end up costing $3-5 per western blot which is 90% of the > > cost of the whole procedure. > > > > People must have some good homemade HRP (and AlkPhos) Chemiluminescent > > Detection protocols or references? > > > > Could you send me one or point me to it? Thanks. > > *Luminol 4mg/ml in DMSO 1ml > *p-iodophenol 1mg/ml in DMSO 1ml > 1M Tris.HCL pH 7.5 0.6ml > H2O2 (30%) 5 microL > H2O 7.5ml > > *Store at -20 > Mix this stuff up fresh a few minutes before use. > Works for me! Where can I buy the luminol and the p-iodophenol from Thanks in advance Ali From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!agate!spool.mu.edu!uwm.edu!www.nntp.primenet.com!nntp.primenet.com!news.maxwell.syr.edu!news-feed.inet.tele.dk!enews.sgi.com!news.corp.sgi.com!news.sgi.com!csulb.edu!info.ucla.edu!nnrp.info.ucla.edu!usenet From: Edmundo Castro Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Substrates? Date: Mon, 02 Jun 1997 09:20:19 -0700 Organization: UCLA Lines: 3 Message-ID: <3392F2C3.7E7C@physci.ucla.edu> References: <338EFCA7.41C6@risotto.mit.edu> <338ECA5B.6690@uic.edu> Reply-To: ecastro@physci.ucla.edu NNTP-Posting-Host: 149.142.241.23 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01 (Macintosh; I; PPC) To: Keld Sorensen Xref: biosci bionet.cellbiol:7417 bionet.immunology:11890 bionet.molbio.methds-reagnts:58198 bionet.molbio.proteins:10888 I have reused primary Ab >15 times with good results. Save $ =) EC From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!uwm.edu!msunews!netnews.upenn.edu!mail.med.upenn.edu!rcowherd From: rcowherd@mail.med.upenn.edu (Robert Cowherd) Newsgroups: bionet.molbio.proteins Subject: Large expression from rare mrna? Date: 2 Jun 1997 15:41:16 GMT Organization: University of Pennsylvania Lines: 16 Message-ID: <5mupis$mrj@netnews.upenn.edu> NNTP-Posting-Host: mail.med.upenn.edu X-Newsreader: TIN [version 1.2 PL2-upenn1.1] Can any one give me an example or two of a moderate to large amount of protein while the steady state of the message is low? I have a proimflammatory cytokine which generates 1600-3000 pg/ml by elisa but seems to be undetectable by northern. Boss wants other examples. Anyone have the? Or how about alternate hypothesis to explain these results? Any help is appreciated. Thank you, Bob Cowherd UPENN (215) 662-2071 From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!GATE.SINICA.EDU.TW!bcschiou From: bcschiou@GATE.SINICA.EDU.TW (Shean-Tai Chiou) Newsgroups: bionet.molbio.proteins Subject: Re: Tissue tearor - which one? Date: 2 Jun 1997 09:02:05 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <5mubc3$1jr$1@surz03fi.HRZ.Uni-Marburg.DE> NNTP-Posting-Host: net.bio.net You can get it from Scienceware. S.T. Chiou IBCAS On 2 Jun 1997, Hasse Christof wrote: > Hi everybody out in netland! > > I'm looking for an tissue homogenizer which can be used in > 1.5ml Eppendorf cups. Do You know some? > > Thank You very much! > > > chris > > From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!www.nntp.primenet.com!nntp.primenet.com!EU.net!feeder.chicago.cic.net!chi-news.cic.net!ftpbox.mot.com!newsfeed.acns.nwu.edu!news.cc.uic.edu!not-for-mail From: Keld Sorensen Newsgroups: bionet.molbio.proteins Subject: Re: Tissue homogenizer for Eppies? Date: Mon, 02 Jun 1997 08:48:31 +0000 Organization: University of Illinois, College of Medicine Lines: 3 Message-ID: <339288DD.27C9@uic.edu> References: <5mubc3$1jr$1@surz03fi.HRZ.Uni-Marburg.DE> NNTP-Posting-Host: slip-rock-02.dialin.uic.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01 (Macintosh; I; PPC) To: Hasse Christof I think Kontes sell these with a little motorized pestle. Keld. From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 From: "Dr E. Buxbaum" Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Substrates? Date: 2 Jun 1997 13:07:54 GMT Organization: University of Leicester (PCFS User) Lines: 10 Message-ID: <5mugja$1ds@falcon.le.ac.uk> References: <338EFCA7.41C6@risotto.mit.edu> <338ECA5B.6690@uic.edu> NNTP-Posting-Host: pc47.msb.le.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) Path: biosci!rutgers.rutgers.edu!gatech!news1.mid-ga.com!nntp.mid-ga.com!news.oru.edu!news-spur1.maxwell.syr.edu!news.maxwell.syr.edu!rill.news.pipex.net!pipex!server1.netnews.ja.net!warwick!leicester!usenet Xref: biosci bionet.cellbiol:7415 bionet.immunology:11885 bionet.molbio.methds-reagnts:58186 bionet.molbio.proteins:10884 Keld Sorensen wrote: >Say you use 10 ml of a 1:500 dilution of a primary antibody that cost >you $1,000/mg, then the antibody cost is $20 per blot. If you had to use >say a 1:250 dilution (weak affinity antibody) then the cost would be $40 >per blot!! This antibody solution would be reused many, many times. The costs per blot are, therefore, much lower. From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!agate!hammer.uoregon.edu!newsfeed.internetmci.com!news-was.dfn.de!news-fra1.dfn.de!zeus.rbi.informatik.uni-frankfurt.de!grapool30.rz.uni-frankfurt.de!News.Uni-Marburg.DE!not-for-mail From: hassec@Mailer.Uni-Marburg.DE (Hasse Christof) Newsgroups: bionet.molbio.proteins Subject: Tissue tearor - which one? Date: 2 Jun 1997 11:38:43 GMT Organization: HRZ Uni Marburg Lines: 9 Message-ID: <5mubc3$1jr$1@surz03fi.HRZ.Uni-Marburg.DE> NNTP-Posting-Host: pprz03.hrz.uni-marburg.de X-Newsreader: TIN [UNIX 1.3 BETA-950824-color PL0] Hi everybody out in netland! I'm looking for an tissue homogenizer which can be used in 1.5ml Eppendorf cups. Do You know some? Thank You very much! chris From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!howland.erols.net!news.maxwell.syr.edu!nntp.uio.no!news.kth.se!news.datakom.su.se!not-for-mail From: Stefan Backstrom Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Electroporation of S. cerevisiae Date: Mon, 02 Jun 1997 13:29:13 +0200 Organization: Stockholm University Lines: 22 Message-ID: <3392AE89.41C6@alfa.mbb.ki.se> NNTP-Posting-Host: zeta.mbb.ki.se Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (X11; I; IRIX 5.3 IP22) Xref: biosci bionet.molbio.methds-reagnts:58182 bionet.molbio.proteins:10882 Hello there. I'm looking for the Protocol with big P for the electroporation of S. cerevisiae cells. Can you help me out ?? We also use a homemade chemiluminescent coctail. Works as good as Amershans and slighter better than p-iodophenol. Modified according to A. Diaz et al, J BIOLUMIN CHEMILUMIN 1995; 10: 175-84 Solution A 68 mM p-coumaric acid (Mw 164.2, C-9008, Sigma) in DMSO. Store at 100 microlitre aliquots at -20 Solution B 1.25 mM luminol(Mw 177.2, A8511, Sigma) in 0.1 M tris, pH 8.5. First dissolve luminol in a small volume of NaOH and then dilute with 0.1 M tris, pH 8.5. Store in aliquots of 10 ml at -20. Mix one A with one B and add 30 microlitres of 3 % hydrogen peroxide. Incubate the blot for one minute and immediately wrap it in plastic foil and expose for 30 s, 1 and 5 minutes. From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!agate!nntpfeed.doc.ic.ac.uk!sunsite.doc.ic.ac.uk!nntp0.brunel.ac.uk!strath-cs!news.qub.ac.uk!not-for-mail From: Andrew Wallace Newsgroups: bionet.molbio.proteins Subject: Re: peptide synthesis Date: Mon, 02 Jun 1997 09:51:10 +0100 Organization: Queens University Belfast Lines: 32 Message-ID: <3392897D.634A@qub.ac.uk.see.signature> References: Reply-To: a.wallace@qub.ac.uk.see.signature NNTP-Posting-Host: awall.bc.qub.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (Macintosh; I; PPC) Rob wrote: > > Hi > Does anyone know of a way to incorporate a label (e.g. biotin or > fluorescent tag) into a peptide during solid-phase synthesis? - I suppose > I'm really asking if labelled protected amino acids are commercially > available... > Thanks in advance > Rob Hi Rob, It's fairly easy to incorporate biotin and a number of fluorescent labels as the final residue of the N-terminal of the peptide during synthesis. Biotin has a free carboxyl group which can be activated by conventional uronium salt (HBTU, etc.) activation methods; it also survives the acid cleavage and workup procedures (at least, it does in Fmoc-polyamide chemistry with TFA - I haven't tried HF cleavage). If you want to label the side chains of your peptide you either need to prepare the appropriate labelled protected amino acid beforehand or choose your side chain protecting groups carefully to allow selective deprotection during synthesis. There are many possibilities, but any competent peptide lab should be able to help you. Andrew -- - note antispam feature in return address. My real address is: ================================================================== Andrew Wallace,Ph.D., Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html ================================================================== From owner-proteins@net.bio.net Sun Jun 01 23:00:00 1997 Path: biosci!agate!spool.mu.edu!uwm.edu!chi-news.cic.net!su-news-hub1.bbnplanet.com!su-news-feed4.bbnplanet.com!news.bbnplanet.com!enews.sgi.com!news.corp.sgi.com!news.sgi.com!csulb.edu!not-for-mail From: dngo@engr.csulb.edu (Duong Ngo) Newsgroups: bionet.molbio.proteins Subject: Please help!!! Date: 2 Jun 1997 07:12:02 GMT Organization: Cal State Long Beach Lines: 3 Message-ID: <5mtro2$5hq@hatathli.csulb.edu> NNTP-Posting-Host: heart.engr.csulb.edu X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Does anyone know the chemical formulas for alcohol dehydrogenase and hemoglobin or their molecular weights? Please let me know. Thanks in advance! From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Path: biosci!agate!hammer.uoregon.edu!news-xfer.netaxs.com!news.maxwell.syr.edu!newsfeed.internetmci.com!swrinde!rill.news.pipex.net!pipex!server1.netnews.ja.net!warwick!leicester!usenet From: "Dr E. Buxbaum" Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: 3 Jun 1997 12:26:37 GMT Organization: University of Leicester (PCFS User) Lines: 10 Message-ID: <5n12ht$prs@falcon.le.ac.uk> References: <338EFCA7.41C6@risotto.mit.edu> <339318A0.270E@bilbo.bio.purdue.edu> NNTP-Posting-Host: pc47.msb.le.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) Xref: biosci bionet.cellbiol:7423 bionet.immunology:11897 bionet.molbio.methds-reagnts:58227 bionet.molbio.proteins:10893 Ali McBride wrote: >Where can I buy the luminol and the p-iodophenol from >Thanks in advance >Ali 4-Iodophenol is available from Aldrich, Luminol from Fluka. Warning: Do not buy Luminol from Sigma, we had a complete failure with that stuff once. From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Newsgroups: bionet.molbio.proteins Path: biosci!agate!spool.mu.edu!uwm.edu!newsfeeds.sol.net!europa.clark.net!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!btnet!server2.netnews.ja.net!server3.netnews.ja.net!server4.netnews.ja.net!server5.netnews.ja.net!server6.netnews.ja.net!str-ccsun!strath-cs!liv!news From: lgbell@liverpool.ac.uk Subject: Re: peptide synthesis Content-Type: text/plain; charset=us-ascii Message-ID: <3393EFA5.741@liverpool.ac.uk> Sender: news@liverpool.ac.uk (News System) Nntp-Posting-Host: pc028010.med.liv.ac.uk Content-Transfer-Encoding: 7bit Organization: The University of Liverpool References: Mime-Version: 1.0 Date: Tue, 3 Jun 1997 10:19:17 GMT X-Mailer: Mozilla 2.02 (Win16; I) Lines: 29 Rob wrote: > > Hi > Does anyone know of a way to incorporate a label (e.g. biotin or > fluorescent tag) into a peptide during solid-phase synthesis? - I suppose > I'm really asking if labelled protected amino acids are commercially > available... > Thanks in advance > Rob I havent seen any advertised or in the catalogues when buying normal protected amino acids. But really it depends where you want your label as to how you procede. At the N terminus: then you could couple it in a simmilar manner to an amino acid. At the C terminus: Cleave the protected peptide and then couple this to the label by solution phase chemistry before a final deprotection of the N terminus and side chains. In the middle of the chain: perhaps allyl protected amino acids, which are commercially available (I think Nova biochem amongst others supply them), could be used. Making the peptide as normal with the appropriate residue to be labelled coupled as its allyl derivative. At the end of the synthesis selectively remove the allyl group and couple you label. Then cleave as normal. Hope this helpas, Len Bell. lgbell@liv.ac.uk From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Path: biosci!agate!usenet.INS.CWRU.Edu!gatech!howland.erols.net!ix.netcom.com!news From: hk-miami@ix.netcom.com(HK) Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: 4 Jun 1997 01:02:30 GMT Organization: Netcom Lines: 22 Message-ID: <5n2er6$n0m@dfw-ixnews8.ix.netcom.com> References: <338EFCA7.41C6@risotto.mit.edu> <339318A0.270E@bilbo.bio.purdue.edu> <5n12ht$prs@falcon.le.ac.uk> NNTP-Posting-Host: mia-fl13-11.ix.netcom.com X-NETCOM-Date: Tue Jun 03 8:02:30 PM CDT 1997 Xref: biosci bionet.cellbiol:7430 bionet.immunology:11903 bionet.molbio.methds-reagnts:58254 bionet.molbio.proteins:10899 x-no-archive: yes In <5n12ht$prs@falcon.le.ac.uk> "Dr E. Buxbaum" writes: >Ali McBride wrote: >>Where can I buy the luminol and the p-iodophenol from >>Thanks in advance >>Ali >------------------------------------- >4-Iodophenol is available from Aldrich, Luminol from Fluka. Warning: >Do not buy Luminol from Sigma, we had a complete failure with that stuff once. ---------------------------------------------- Maybe it wasn't the luminol...Maybe you mixed or measured something incorrectly, since you said that you only tried it once. I buy Luminol from Sigma...it works just fine. In a side-by-side test of my solutions and the ones in the Amersham ECL kit, my solutions work equally as well. Helene From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Path: biosci!agate!newsfeed.kornet.nm.kr!news.maxwell.syr.edu!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!newsfeed.nacamar.de!fu-berlin.de!news-ber1.dfn.de!news-ham1.dfn.de!news-han1.dfn.de!newsserver.rrzn.uni-hannover.de!infosrv.rz.uni-kiel.de!not-for-mail From: Sibylle Nold Newsgroups: bionet.molbio.proteins Subject: Re: Please help!!! Date: Wed, 04 Jun 1997 01:03:49 -0700 Organization: Rechenzentrum der Universitaet Kiel, Germany Lines: 8 Message-ID: <33952165.63CA@mail.uni-kiel.d400.de> References: <5mtro2$5hq@hatathli.csulb.edu> NNTP-Posting-Host: 134.245.99.226 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01 [de] (Win16; I) Duong Ngo wrote: > > Does anyone know the chemical formulas for alcohol dehydrogenase and > hemoglobin or their molecular weights? Please let me know. Thanks in > advance! I happened to find the molecular weight of hemoglobin, 64 500, hope that helps. From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!howland.erols.net!feed1.news.erols.com!news-xfer.netaxs.com!netnews.upenn.edu!dsinc!newsfeed.pitt.edu!pelli.pathology.pitt.edu!user From: pxpst2+@pitt.edu (peter) Newsgroups: bionet.molbio.proteins Subject: Re: I need a book Date: 3 Jun 1997 20:08:28 GMT Organization: Univ.of Pittsburgh Lines: 11 Message-ID: References: <01bc6b9f$afaf3ac0$0ac8c8c0@jones> NNTP-Posting-Host: pelli.pathology.pitt.edu In article <01bc6b9f$afaf3ac0$0ac8c8c0@jones>, "Patrick R. Jones" wrote: Maniatis is a three volume book covering many of those topics but the seperation sciences are to themselves. Check with the local college and grap an analytical chem book . This will cover the seperation techniques and the princibles to most of the biological techniques Peter Pediaditakis pxpst2@vms.cis.pitt.edu From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Path: biosci!agate!spool.mu.edu!uwm.edu!chi-news.cic.net!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!news.ums.edu!haven.umd.edu!news.umbc.edu!kcarter From: kcarter@umbc.edu (Mr. Ken Carter) Newsgroups: bionet.molbio.proteins Subject: Final Announcement: Extremophile Workshop Date: 3 Jun 1997 19:13:56 GMT Organization: University of Maryland, Baltimore County Lines: 64 Message-ID: <5n1qdk$961@news.umbc.edu> NNTP-Posting-Host: f-umbc7.umbc.edu NNTP-Posting-User: kcarter X-Newsreader: TIN [version 1.2 PL2] THEORY AND TECHNIQUES FOR EXTREMOPHILE RESEARCH JULY 7-10, 1997 Center of Marine Biotechnology (COMB) , Baltimore, Maryland Co-sponsored by American Type Culture Collection (ATCC) and the Maryland Biotechnology Institute (MBI) This four-day laboratory-intensive course will cover procedures, methodologies and up-to-date information about the Archaea, a group of organisms commonly found in what are considered harsh or extreme environments. This workshop provides introductory and intermediate level instruction in Archaeal laboratory techniques, including growth, biochemistry, and genetics. A basic knowledge of microbiology and nucleic acid laboratory procedures is helpful. The first day of the course will be devoted to a symposium on topics in ARCHAEAL research with Methanogenic,(Hyper)Thermophilic and Halophilic Archaea, including: molecular biology, genetics and physiology, ecology, phylogeny, genome organization, genomic applications, consortia and bioremediation, virology, archaeal lipids and lipid biochemistry, and biotechnological applications. The following three days will be devoted to laboratory sessions, which will be preceded by a lecture on methodology and procedures for each of the Archaeal groups. The laboratory sessions will include three two-hour blocks of demonstration and hands-on experiments focusing on Methanogenic, Thermophilic, and Halophilic Archaea and includes media preparation, inoculation, growth, plating, counting, anaerobic biochemistry, transformation, virus induction, and transfection. The broad hands-on experience should give attendees the necessary background and information to conduct similar studies in their own laboratories. This workshop will benefit those with a foreseeable need to use Archaeal systems in their studies. "Archaea: A Laboratory Manual," published by Cold Spring Harbor Laboratory Press, will be used as a framework for the experiments conducted in the workshop. Registration information and a complete Workshop Schedule are available in the WWW at: http://www.atcc.org/workshops/arch_sch.html PLEASE NOTE: This workshop will be conducted at the University of Maryland's Center of Marine Biotechnology (COMB) in the Columbus Center, located in Baltimore's Inner Harbor. Faculty: Harold J. Schreier, Ph.D., University of Maryland Biotechnology Institute (Workshop Co-director); Kevin R. Sowers, Ph.D., UMBI (Workshop Co-director); Frank T. Robb, Ph.D., UMBI; Alan A. Place, Ph.D., UMBI; Shiladitya DasSarma, Ph.D., Univ. Of Massachusetts, Amherst FEEs: $195.00 - One day symposium - limited to 65 participants / 0.7 CEUs $1,195.00 - Symposium and a 3-day laboratory workshop - limited to 24 participants / 2.8 CEUs -- ****************************************************************************** Ken Carter ^ American Type Culture Collection / l \ (301) 231-5525 12301 Parklawn Drive /___l___\__ fax:(301) 816-4364 Rockville, Maryland 20852 \______/ kcarter@atcc.org ~~~~~~~~~~~~~~ From owner-proteins@net.bio.net Mon Jun 02 23:00:00 1997 Path: biosci!ihnp4.ucsd.edu!munnari.OZ.AU!news.ecn.uoknor.edu!feed1.news.erols.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!rill.news.pipex.net!pipex!server1.netnews.ja.net!warwick!news.nott.ac.uk!Fergus.Doherty From: Fergus.Doherty@nottingham.ac.uk (Fergus Doherty) Newsgroups: bionet.molbio.proteins Subject: Acid ppt of proteins? Date: Mon, 02 Jun 1997 16:51:40 +0100 Organization: Nottingham University Lines: 15 Message-ID: NNTP-Posting-Host: wmbpm8.nottingham.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Newsreader: Yet Another NewsWatcher 2.4.0 When I do pulse-chase expts in yeast and ppt proteins with 10% TCA, even at zero time in the chase (after washes to chase out the label), I still get high counts in the TCA soluble supernatant. Any clues? I was wondering if this could be small peptides (but don't know). If so what could I use to ppt them? Higher TCA conc? Perchloric acid? some time bak I seem to remember phosphotungstic acid mentioned, which will (I think) ppt peptides. Any sensible suggestion welcomed. -- Fergus Doherty, Dept Biochemistry, Nottingham University, Fergus.Doherty@nottingham.ac.uk 0115 970 9366 (74-41366 internal) From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!uwm.edu!feeder.chicago.cic.net!news.maxwell.syr.edu!feed1.news.erols.com!newsfeed.internetmci.com!jump.net!grunt.dejanews.com!not-for-mail Date: Wed, 04 Jun 1997 03:52:44 -0600 From: tthamm@gwdg.de Subject: transcription factor HNF-4 Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.yeast,bionet.molbio.proteins Message-ID: <865413980.3570@dejanews.com> Organization: University of Goettingen, Germany; Division: Human Genetics X-Article-Creation-Date: Wed Jun 04 08:46:20 1997 GMT X-Originating-IP-Addr: 134.76.134.159 (unihg9.ukhg.gwdg.de) X-Http-User-Agent: Mozilla/2.0 (Win16; I) X-Authenticated-Sender: tthamm@gwdg.de Lines: 12 Xref: biosci bionet.molbio.methds-reagnts:58265 bionet.molbio.yeast:7158 bionet.molbio.proteins:10900 Hi netters! Has anybody out there antibodys against the transcription factor HNF-4 ? It is a member of the steroid hormone receptor superfamily. Thanks Tarvo -------------------==== Posted via Deja News ====----------------------- http://www.dejanews.com/ Search, Read, Post to Usenet From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!BOTANY.UFL.EDU!jshao From: jshao@BOTANY.UFL.EDU Newsgroups: bionet.molbio.proteins Subject: Double-stranded RF M13 preparation Date: 4 Jun 1997 12:30:19 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hi, I'm trying to prepare the double-stranded replication form of M13 vector for a library construction. I used Wizard Maxiprep system for the purification. The vector size is 8.35kb. By runnig 0.8% agarose gel, I got two bands M.W. close to 20kb and 7kb (major band) which ran at about 8kb as a single band after digestion. The problem is I found up to 10 additional bands with M.W. below 4 kb to 0.5 kb, which persisted after digestion, and the band pattern didn't change. I want to know what are these extra bands and how I can get rid of them. I appreciate your help. Jiahong Shao Dept. of Botany Univ. of Fl From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Larry Hunter Newsgroups: bionet.molbio.proteins,news.announce.conferences Subject: Second Call for Papers, 1998 Pacific Symposium on Biocomputing Date: 4 Jun 1997 10:40:47 -0700 Organization: National Library of Medicine Lines: 318 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net REMINDER: PAPERS ARE DUE JULY 14! CHECK OUR WEB PAGE FOR SESSION DETAILS... >>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>> Call For Papers, Abstracts and Demonstrations ^ v ^ for the v ^ v ^ Pacific Symposium on Biocomputing v ^ v ^ Kapalua, Maui (Hawaii) - January 4-9, 1998 v ^ v URL: http://www.cgl.ucsf.edu/psb <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<< The third Pacific Symposium on Biocomputing (PSB), will be held January 4-9, 1998 in Maui, Hawaii. PSB will bring together top researchers from North America, the Asian Pacific nations, Europe and around the world to exchange research results and address open issues in all aspects of computational biology. PSB will provide a forum for the presentation of work in databases, algorithms, interfaces, visualization, modeling and other computational methods, as applied to biological problems, with emphasis on applications in data-rich areas of molecular biology. PSB intends to attract a balanced combination of computer scientists and biologists, presenting significant original research, demonstrating computer systems, and facilitating formal and informal discussions on topics of importance to computational biology. To provide focus for the very broad area of biological computing, PSB is organized into a series of specific sessions. Each session will involve both formal research presentations and open discussion groups. The 1998 PSB sessions are: * Gene Expression and Genetic Networks * Molecules to Maps: Tools for Visualization & Interaction * Gene Structure Identification in Large-scale Genomic Sequence * Molecular Modeling in Drug Design and Biotechnology * Protein Structure Prediction * The Relationship Between Protein Structure and Function * Computing with Biomolecules * Complexity and Information Theoretic Approaches to Biology * Distributed and Intelligent Databases * Building Bioinformation Infrastructure in the Pacific Rim Further details on each session can be found below. The core of the conference consists of rigorously peer-reviewed full-length papers reporting on original work. Accepted papers will be published in a hard-bound archival proceedings, and the best of these will be presented orally to the entire conference. Researchers wishing to present their research without official publication are encouraged to submit a one page abstract, and present their work in discussion, poster and demonstration sessions. Important dates: Paper submissions due: July 14, 1997 Notification of paper acceptance: August 22, 1997 Final paper deadline September 22, 1997 Abstract deadline October 1, 1997 Meeting January 4-9, 1998 Paper format: Papers may be up to 12 single spaced pages, and *must* use our supplied format, available from ftp://ftp-smi.stanford.edu/pub/altman/psb. Each paper must be accompanied by a cover letter stating that it contains original unpublished results not currently under consideration elsewhere and that all co-authors concur with its contents. Please indicate in your cover letter for which specific session (if any) you wish your paper or abstract to be considered. Papers and abstracts may be submitted electronically. Contact Russ Altman (russ.altman@stanford.edu) for additional information. Paper submission address: For physical submission, please send five copies of your paper to: PSB-98 c/o Russ B. Altman Section on Medical Informatics SUMC, MSOB X-215 Stanford, CA, USA 94305-5479 (415) 725-0659 Electronic submission of papers and abstracts is encouraged. Contact Dr. Altman for information about electronic submission. Travel support: We have been able to offer partial travel support to many PSB attendees in the past, including most authors of accepted full papers who request support. However, due to our sponsoring agencies' schedules, we are unable to offer travel awards before the registration (and payment) deadlines for authors. We recognize that this is inconvenient, and we are doing our best to rectify the situation. NO ONE IS GUARANTEED TRAVEL SUPPORT. Travel support applications will be available on our web site. Conference cochairs: Russ Altman, Stanford University A. Keith Dunker, Washington State University Lawrence Hunter, National Library of Medicine Teri Klein, University of California, San Francisco Each session has a chair who is responsible for organizing submissions. Please contact the specific session chair relevant to your interests for further information. PSB '98 Sessions: *** Gene Expression and Genetic Networks Cochairs: Barbara Bryant, Aleksandar Milosavljevic & Roland Somogyi Computational methods in the monitoring, analysis, and modeling of RNA and protein expression; gene regulatory network models and new methods of acquiring and analyzing large-scale gene expression data. Contact: Roland Somogyi Phone: +1 (301) 402-1407 Fax: +1 (301) 402-1565 Email: rolands@helix.nih.gov Web: http://www.cgl.ucsf.edu/psb/sessions/expression.html *** Molecules to Maps: Tools for Visualization & Interaction Cochairs: Tom Ferrin & Eileen Kraemer Tools and techniques to assist scientists in evaluating, absorbing, navigating, and correlating sequence, structural, and functional data through visualization and user interaction. Contact: Eileen Kraemer Phone: +1 (314) 935-6621 Fax: +1 (314) 935-7302 Email: eileen@cs.wustl.edu Web: http://www.cgl.ucsf.edu/psb/sessions/visualization.html *** Gene Structure Identification in Large-scale Genomic Sequence Cochairs: Ying Xu, Edward Uberbacher Any aspect of computational gene finding, particularly how to fully utilize the available EST/protein sequences and biological information, statistical and mathematical tools to automate gene identification and annotation in large-scale genomic sequences. Contact: Ying Xu Phone: +1 (423) 574-7263 Fax: +1 (423) 574-7860 Email: xyn@@ornl.gov Web: http://www.cgl.ucsf.edu/psb/sessions/gene.html *** Molecular Modeling in Drug Design and Biotechnology Cochairs: Terry Lybrand, Teri Klein, Jurgen Bajorath State-of-the-art molecular modeling approaches which aid in small molecular and structure-based drug design and protein engineering. Contact: Terry Lybrand Phone: +1 (206) 685-1515 Fax: +1 (206) 616-4387 E-mail: lybrand@proteus.bioeng.washington.edu Web: http://www.cgl.ucsf.edu/psb/sessions/modeling.html *** Protein Structure Prediction Chair: Richard Lathrop All aspects of protein structure prediction, with emphasis on approaches that lead to testable protein structure predictions, and experimental results across a large diverse set of proteins. Contact: Richard Lathrop Phone: +1 (714) 824-4021 Fax: +1 (714) 824-4056 Email: rickl@ics.uci.edu Web: http://www.cgl.ucsf.edu/psb/sessions/psp.html *** The Relationship Between Protein Structure and Function: How Have Proteins Over Time Diverged in Function? Cochairs: Patricia Babbitt and Monica Riley Computational strategies to address the "structure-function" problem, particularly the interface between automated structural analysis, evolutionary change and biological insight. Contact: Patricia Babbitt Phone: +1 (415) 476-3784 Fax: +1 (415) 476-0688 email: babbitt@cgl.ucsf.edu web: http://www.cgl.ucsf.edu/psb/sessions/function.html *** Computing with Biomolecules Cochairs: Peter Clote, Masami Hagiya, Tom Head Both artificial and naturally occurring computations in which biological macromolecules act as computational elements. Contact: Tom Head Phone: +1 (607) 777-2278 Fax: +1 (607) 777-2450 Email: Tom@Math.Binghamton.edu Web: http://www.cgl.ucsf.edu/psb/sessions/compute.html *** Complexity and Information Theoretic Approaches to Biology Cochairs: David Dowe and Klaus Prank Approaches to biological problems using notions of information or complexity, including methods such as Algorithmic Probability, Minimum Message Length and Minimum Description Length. Two possible applications are (e.g.) protein folding and biological information processing. Contact: David Dowe Phone: +61 3 9905-5776 Fax: +61 3 9905-5146 Email: dld@cs.monash.edu.au Web: http://www.cgl.ucsf.edu/psb/sessions/info.html *** Distributed and Intelligent Databases Cochairs: Dmitrij Frishman and Patrick Argos Computer and algorithmic methods that result in more intelligent, interconnected and accessible molecular biological databases. Contact: Dmitrij Frishman Phone: +49 (89) 8578-2664 Fax: +49 (89) 8578-2655 Email: frishman@mips.biochem.mpg.de Web: http://www.cgl.ucsf.edu/psb/sessions/database.html *** Building Bioinformation Infrastructure in the Pacific Rim Cochairs: S. Subbiah, T.W. Tan, Tim Littlejohn, Hideaki Sugawara Collaboration and cooperation to create a shared biological information infrastructure across the Pacific Rim nations and beyond, that will guarantee a quality of service to the users of our biocomputing and bioinformatics resources. Of particular emphasis in this session is how to transfer the technology to research organizations in developing nations which find difficulty in accessing biocomputing and bioinformatics services. Contact: Tin Wee Tan Phone: +65 772-6490 Fax: +65 872-6205 Email: tinwee@bic.nus.sg Web: http://www.cgl.ucsf.edu/psb/sessions/pacific.html For further information about the conference, registration, possible travel support, submission of papers not covered by the above categories, or other information, please contact the conference coordinator: Norma Belfer PSB Conference Coordinator UCSF Computer Graphics Laboratory Box 0446 513 Parnassus Avenue San Francisco, California 94143-0446 email: psb@cgl.ucsf.edu fax: +1 (415) 476-0688 tel: +1 (415) 476-5128 -- Lawrence Hunter, PhD. National Library of Medicine phone: +1 (301) 496-9303 Bldg. 38A, 9th fl, MS-54 fax: +1 (301) 496-0673 Bethesda. MD 20894 USA email: hunter@nlm.nih.gov From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!daresbury!server5.netnews.ja.net!server3.netnews.ja.net!nntpfeed.doc.ic.ac.uk!sunsite.doc.ic.ac.uk!warwick!leicester!usenet From: "Dr E. Buxbaum" Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: 4 Jun 1997 17:16:40 GMT Organization: University of Leicester (PCFS User) Lines: 13 Message-ID: <5n47to$188@falcon.le.ac.uk> References: <338EFCA7.41C6@risotto.mit.edu> <339318A0.270E@bilbo.bio.purdue.edu> <5n12ht$prs@falcon.le.ac.uk> <5n2er6$n0m@dfw-ixnews8.ix.netcom.com> NNTP-Posting-Host: pc47.msb.le.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) Xref: biosci bionet.cellbiol:7438 bionet.immunology:11913 bionet.molbio.methds-reagnts:58283 bionet.molbio.proteins:10903 hk-miami@ix.netcom.com(HK) wrote: >Maybe it wasn't the luminol...Maybe you mixed or measured something >incorrectly, since you said that you only tried it once. >I buy Luminol from Sigma...it works just fine. In a side-by-side >test of my solutions and the ones in the Amersham ECL kit, my >solutions work equally as well. I would not say things like this if I hadn't done the ovious tests. In general I had quite a few problems with Sigma's chemicals over the years, for example SDS which was insoluble in water, inactive enzyme... Looks like a quality control problem to me. From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!www.nntp.primenet.com!nntp.primenet.com!europa.clark.net!cpk-news-hub1.bbnplanet.com!cam-news-feed2.bbnplanet.com!news.bbnplanet.com!dilbert.whoi.edu!news@whoi.edu From: Eli Hestermann Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: Wed, 04 Jun 1997 10:16:40 -0400 Organization: Woods Hole Oceanographic Institution Lines: 22 Message-ID: <339578C8.5F2C@whoi.edu> References: <338EFCA7.41C6@risotto.mit.edu> Reply-To: ehestermann@whoi.edu NNTP-Posting-Host: octavian.whoi.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01 (Win95; I) Xref: biosci bionet.cellbiol:7433 bionet.immunology:11910 bionet.molbio.methds-reagnts:58274 bionet.molbio.proteins:10902 David Aldridge wrote: > *Luminol 4mg/ml in DMSO 1ml > *p-iodophenol 1mg/ml in DMSO 1ml > 1M Tris.HCL pH 7.5 0.6ml > H2O2 (30%) 5 microL > H2O 7.5ml > > *Store at -20 > Mix this stuff up fresh a few minutes before use. > Works for me! I assume this is for HRP, does it work for AP, too? If not, does anybody have one for AP? Eli Hestermann -- Eli V. Hestermann ehestermann@whoi.edu Woods Hole Oceanographic Institution Massachusetts Institute of Technology "Vita brevis est, ars longa" - Seneca From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!howland.erols.net!newsxfer.itd.umich.edu!yale!oitnews.harvard.edu!das-news2.harvard.edu!goldenapple.srv.cs.cmu.edu!bb3.andrew.cmu.edu!newsfeed.pitt.edu!pelli.pathology.pitt.edu!user From: pxpst2+@pitt.edu (peter) Newsgroups: bionet.molbio.proteins Subject: Re: Please help!!! Date: 3 Jun 1997 20:10:24 GMT Organization: Univ.of Pittsburgh Lines: 12 Message-ID: References: <5mtro2$5hq@hatathli.csulb.edu> NNTP-Posting-Host: pelli.pathology.pitt.edu In article <5mtro2$5hq@hatathli.csulb.edu>, dngo@engr.csulb.edu (Duong Ngo) wrote: > Does anyone know the chemical formulas for alcohol dehydrogenase and > hemoglobin or their molecular weights? Please let me know. Thanks in > advance! proteins can be easily found in protein database online or in the library. Peter Pediaditakis From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 From: hk-miami@ix.netcom.com(HK) Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: 5 Jun 1997 00:01:15 GMT Organization: Netcom Lines: 25 Message-ID: <5n4vkb$st0@sjx-ixn4.ix.netcom.com> References: <338EFCA7.41C6@risotto.mit.edu> <339318A0.270E@bilbo.bio.purdue.edu> <5n12ht$prs@falcon.le.ac.uk> <5n2er6$n0m@dfw-ixnews8.ix.netcom.com> <5n47to$188@falcon.le.ac.uk> NNTP-Posting-Host: mia-fl4-11.ix.netcom.com X-NETCOM-Date: Wed Jun 04 5:01:15 PM PDT 1997 Path: biosci!rutgers.rutgers.edu!gatech!news1.mid-ga.com!news.hom.net!nntp.mid-ga.com!news.oru.edu!news-spur1.maxwell.syr.edu!news.maxwell.syr.edu!ix.netcom.com!news Xref: biosci bionet.cellbiol:7441 bionet.immunology:11918 bionet.molbio.methds-reagnts:58293 bionet.molbio.proteins:10907 x-no-archive: yes In <5n47to$188@falcon.le.ac.uk> "Dr E. Buxbaum" writes: > >hk-miami@ix.netcom.com(HK) wrote: > >>Maybe it wasn't the luminol...Maybe you mixed or measured something >>incorrectly, since you said that you only tried it once. >>I buy Luminol from Sigma...it works just fine. In a side-by-side >>test of my solutions and the ones in the Amersham ECL kit, my >>solutions work equally as well. > >I would not say things like this if I hadn't done the ovious tests. In >general I had quite a few problems with Sigma's chemicals over the years, >for example SDS which was insoluble in water, inactive enzyme... Looks >like a quality control problem to me. ----------- I'm not sure what a more oBvious test would be than testing it side by side with the reagents I was trying to duplicate and seeing that both worked equally well. What other test would you suggest? Helene > From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!agate!spool.mu.edu!howland.erols.net!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!news-feed.inet.tele.dk!news.he.net!news.pagesat.net!composer.inav.net!usenet From: Robert J Jensen Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: Wed, 04 Jun 1997 23:14:27 -0500 Organization: Internet Navigator, Inc. Lines: 3 Message-ID: <33963D23.42B2@inav.net> References: <338EFCA7.41C6@risotto.mit.edu> <339318A0.270E@bilbo.bio.purdue.edu> <5n12ht$prs@falcon.le.ac.uk> <5n2er6$n0m@dfw-ixnews8.ix.netcom.com> <5n47to$188@falcon.le.ac.uk> NNTP-Posting-Host: dip109.inav.net Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (Macintosh; I; 68K) Xref: biosci bionet.cellbiol:7443 bionet.immunology:11920 bionet.molbio.methds-reagnts:58299 bionet.molbio.proteins:10908 Over the years I have had very good luck with chemicals from Sigma. In my book they are a top notch comapny. rjj From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!agate!nature.berkeley.edu!lhom From: lhom@nature.berkeley.edu (Louis Hom) Newsgroups: sci.chem,bionet.molbio.proteins,bionet.molbio.methds-reagnts Subject: Reversible covalent-linked protease inhibitors Date: 4 Jun 1997 22:44:59 GMT Organization: University of California, Berkeley Lines: 7 Message-ID: <5n4r5b$i7m@agate.berkeley.edu> NNTP-Posting-Host: nature.berkeley.edu Xref: biosci bionet.molbio.proteins:10906 bionet.molbio.methds-reagnts:58290 Does anyone know of a method to reversibly inhibit cysteine proteases (e.g., cathepsin L) by covalent linkage? Maybe -SS- or Hg? -- _______________________________________________________________________________ Lou Hom >K '93 "No one ever went broke lhom@nature.berkeley.edu underestimating the taste http://www.ocf.berkeley.edu/~lhom of the American public." --Mencken. From owner-proteins@net.bio.net Tue Jun 03 23:00:00 1997 Path: biosci!agate!spool.mu.edu!howland.erols.net!worldnet.att.net!newsadm From: Mark Dalton Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Date: Wed, 04 Jun 1997 23:34:45 -0500 Organization: AT&T WorldNet Services Lines: 22 Message-ID: <339641E5.819@worldnet.att.net> References: <338EFCA7.41C6@risotto.mit.edu> Reply-To: mdalton@worldnet.att.net NNTP-Posting-Host: 207.146.217.179 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.02E (Macintosh; U; PPC) Xref: biosci bionet.cellbiol:7444 bionet.immunology:11923 bionet.molbio.methds-reagnts:58301 bionet.molbio.proteins:10909 I tried this once it was much less sensitive than super signal from Pierce Chemicals. By the way Pierce is 1/2 the price of ECL from Amersham, and is more sensitive. I suggest you try the homemade stuff for yourself but make sure you do the controls the chemicals were very cheap around $20. I found protocol in the Red Book. It is listed in the other replies. Good Luck Mark Thomas Cameron wrote: > > Company kits end up costing $3-5 per western blot which is 90% of the > cost of the whole procedure. > > People must have some good homemade HRP (and AlkPhos) Chemiluminescent > Detection protocols or references? > > Could you send me one or point me to it? Thanks. > > -- > Thomas Cameron From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!biosci!not-for-mail From: newera@plaza.snu.ac.kr Newsgroups: bionet.molbio.proteins, Subject: Acetic acid + NaOH = buffer at pH 5.0? Date: 5 Jun 1997 09:35:21 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: <5n6ps9$2no@net.bio.net> NNTP-Posting-Host: net.bio.net Summary: Keywords: Is a mixture of acetic acid and NaOH a buffer, especially in a concentrated protein solution around 1~5mM? I am sure that 'Acetic acid + Sodium Acetate = buffer at pH 5.0', but not convinced of 'Acetic acid + NaOH = buffer at pH 5.0' because it is not a mixture of weak acid and its salt form. I am definitely in the situation where the latter is much preferred if it can be a buffer at all. Any comment will be appreciated. Best regards, Lee -- Lee, Ji Hyun newera@plaza.snu.ac.kr Laboratory of Physical Pharmacy(Prof. Lee, Bong Jin) College of Pharmacy Seoul National University Shinlim-Dong, Kwanak-Gu Seoul 151-742, Korea. Tel : 82-2-880-7869 Fax : 82-2-872-3632 From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!CC.USU.EDU!arsphys From: arsphys@CC.USU.EDU (Phil Harrison) Newsgroups: bionet.molbio.proteins Subject: Re: Acetic acid + NaOH = buffer at pH 5.0? Date: 5 Jun 1997 12:15:03 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 39 Sender: daemon@net.bio.net Distribution: world Message-ID: <3.0.1.32.19970605132107.006aed04@cc.usu.edu> References: <5n6ps9$2no@net.bio.net> NNTP-Posting-Host: net.bio.net At 09:35 AM 6/5/97 -0700, Lee wrote: >Is a mixture of acetic acid and NaOH a buffer, especially in a concentrated protein solution around 1~5mM? > Protein concentration is probably not important as long as the buffer concentration is 5-10 X greater. >I am sure that 'Acetic acid + Sodium Acetate = buffer at pH 5.0', >but not convinced of 'Acetic acid + NaOH = buffer at pH 5.0' because it is not a mixture of weak acid and its salt form. Acetic acid + NaOH will make a perfectly satisfactory buffer since it is a combination of a *weak* acid and a strong base. If you start with the same molar concentration of acetate (in the form of acetic acid in solution) as you have in the Acetic acid + Sodium Acetate example, then bring the pH to 5.0 with NaOH, you will be creating the same concentration of "Sodium Acetate" (ions) as the other example started with. The results will be identical. The buffering depends on the dissociation constant of acetate, reflected by pKa. Since the pKa for acetate is 4.76, at pH 5.0 you are in the range where acetate buffers well, regardless of how you got there. >I am definitely in the situation where the latter is much preferred if it can be a buffer at all. The only possible advantage I see in using Sodium Acetate + Acetic acid might be to avoid having to add NaOH if the NaOH is not be as pure as the other reagents. That may not even be an issue here. Good luck. I also recommend brushing up on buffers. Phil Harrison From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!daresbury!uninett.no!sn.no!www.nntp.primenet.com!nntp.primenet.com!europa.clark.net!news.maxwell.syr.edu!newsfeed.direct.ca!news.uoregon.edu!news.acsu.buffalo.edu!dsinc!newsfeed.pitt.edu!pelli.pathology.pitt.edu!user From: pxpst2+@pitt.edu (peter) Newsgroups: bionet.molbio.proteins, Subject: Re: Acetic acid + NaOH = buffer at pH 5.0? Date: 5 Jun 1997 17:03:16 GMT Organization: Univ.of Pittsburgh Lines: 32 Distribution: world Message-ID: References: <5n6ptq$3cl@net.bio.net> NNTP-Posting-Host: pelli.pathology.pitt.edu In article <5n6ptq$3cl@net.bio.net>, newera@plaza.snu.ac.kr wrote: .> Summary: .> Keywords: .> .> A mixture of acetic acid and NaOH is a buffer, especially in the case of a .concentrated protein solution around 1~5mM? Protein in solution has nothing to do with whether or not that is a buffer. As for your question in is a buffer system but not a good one because you are using a strong base with a strong acid .> .> I am sure that 'Acetic acid + Sodium Acetate = buffer at pH 5.0', .> but not convinced of 'Acetic acid + NaOH = buffer at pH 5.0' because it is .not a mixture of weak acid and its salt form. To determine the pH one can substitute into the Henderson-Hasselblat equation and solve. Instead of using the term "salt form" call it the conjugate base that is what it is. In solution salts disassociate and are no longer salts. .> .> I am definitely in the situation where the latter is much preferred if it can .be a buffer at all. You should pick up a general chemistry book such as Paulings book and read up you are a little confused with thoughts on buffer systems Peter Pediaditakis pxpst2@vms.cis.pitt.edu From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!csulb.edu!hammer.uoregon.edu!hunter.premier.net!feed1.news.erols.com!howland.erols.net!news.starnet.net!news.starnet.net!newsreader.wustl.edu!swede From: swede@biodec.wustl.edu (Marci Swede) Newsgroups: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Subject: Re: Homemade Chemiluminescent Protocols? Followup-To: bionet.cellbiol,bionet.immunology,bionet.molbio.methds-reagnts,bionet.molbio.proteins, Date: 5 Jun 1997 20:42:19 GMT Organization: Washington University in St. Louis Lines: 32 Message-ID: <5n78bb$kdf$1@newsreader.wustl.edu> References: <338EFCA7.41C6@risotto.mit.edu> <339318A0.270E@bilbo.bio.purdue.edu> <5n12ht$prs@falcon.le.ac.uk> <5n2er6$n0m@dfw-ixnews8.ix.netcom.com> <5n47to$188@falcon.le.ac.uk> <5n4vkb$st0@sjx-ixn NNTP-Posting-Host: biodec.wustl.edu X-Newsreader: TIN [version 1.2 PL0] Xref: biosci bionet.cellbiol:7450 bionet.immunology:11933 bionet.molbio.methds-reagnts:58345 bionet.molbio.proteins:10920 4.ix.netcom.com> Some attributes got messed up here--x the complaint was about Sigma's luminol not working. The request was for testing if the failure was for Sigma luminol or some other reason that batch didn't work. HK (hk-miami@ix.netcom.com) wrote: : x-no-archive: yes : In <5n47to$188@falcon.le.ac.uk> "Dr E. Buxbaum" writes: : > : >hk-miami@ix.netcom.com(HK) wrote: : > : >>Maybe it wasn't the luminol...Maybe you mixed or measured something : >>incorrectly, since you said that you only tried it once. : >>I buy Luminol from Sigma...it works just fine. In a side-by-side : >>test of my solutions and the ones in the Amersham ECL kit, my : >>solutions work equally as well. : > : >I would not say things like this if I hadn't done the ovious tests. In : >general I had quite a few problems with Sigma's chemicals over the : years, : >for example SDS which was insoluble in water, inactive enzyme... Looks : >like a quality control problem to me. : ----------- : I'm not sure what a more oBvious test would be than testing it side by : side with the reagents I was trying to duplicate and seeing that both : worked equally well. What other test would you suggest? : Helene : > From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!csulb.edu!hammer.uoregon.edu!hunter.premier.net!feed1.news.erols.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!baron.netcom.net.uk!netcom.net.uk!nntpfeed.doc.ic.ac.uk!sunsite.doc.ic.ac.uk!charlie.lif.icnet.uk!mac052034.lif.icnet.uk!user From: I.McFarlane@icrf.icnet.uk (Ian McFarlane) Newsgroups: bionet.molbio.proteins Subject: Re: Acid ppt of proteins? Date: Thu, 05 Jun 1997 12:02:41 +0000 Organization: Imperial Cancer Research Fund Message-ID: References: NNTP-Posting-Host: mac052034.lif.icnet.uk X-Newsreader: Value-Added NewsWatcher 2.0b27+ Lines: 33 Hi Fergus, I don't know if you will remember me, I was a postgrad in Frank Hemming's Labs. Whats your label? 35S methionine? Don't most organisms take up amino acids fairly rapidly at t=0 most of the internal label will still be amino acids won't it? As to removal it depends on what you want to look at in the TCA supernatant. But you could try ethanol or acetone precipitations. Ian Mc In article , Fergus.Doherty@nottingham.ac.uk (Fergus Doherty) wrote: > When I do pulse-chase expts in yeast and ppt proteins with 10% TCA, even at > zero time in the chase (after washes to chase out the label), I still get > high counts in the TCA soluble supernatant. Any clues? I was wondering if > this could be small peptides (but don't know). If so what could I use to > ppt them? Higher TCA conc? Perchloric acid? some time bak I seem to > remember phosphotungstic acid mentioned, which will (I think) ppt peptides. > Any sensible suggestion welcomed. > > -- > Fergus Doherty, > Dept Biochemistry, > Nottingham University, > > Fergus.Doherty@nottingham.ac.uk > 0115 970 9366 (74-41366 internal) From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!howland.erols.net!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!baron.netcom.net.uk!netcom.net.uk!nntpfeed.doc.ic.ac.uk!sunsite.doc.ic.ac.uk!nntp0.brunel.ac.uk!strath-cs!news.qub.ac.uk!not-for-mail From: Andrew Wallace Newsgroups: bionet.molbio.proteins Subject: Re: Large expression from rare mrna? Date: Thu, 05 Jun 1997 12:58:48 +0100 Organization: Queens University Belfast Message-ID: <3396A9F7.48E5@qub.ac.uk.see.signature> References: <5mupis$mrj@netnews.upenn.edu> Reply-To: a.wallace@qub.ac.uk.see.signature NNTP-Posting-Host: awall.bc.qub.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (Macintosh; I; PPC) Lines: 35 Robert Cowherd wrote: > > Can any one give me an example or two of a moderate to large amount of > protein while the steady state of the message is low? > > I have a proimflammatory cytokine which generates 1600-3000 pg/ml by > elisa but seems to be undetectable by northern. Boss wants other > examples. Anyone have the? Or how about alternate hypothesis to explain > these results? > > Any help is appreciated. > > Thank you, > > Bob Cowherd > UPENN > (215) 662-2071 Bob, One such candidate may be the various classes of opiate receptor, which are expressed in fairly high amounts (not sure of the quantities) but whose mRNA is almost undetectable. In fact, this was probably one reason that they took nearly 20 years to clone after initial discovery of the proteins. Hope this is of some use, Andrew -- - note antispam feature in return address. My real address is: ================================================================== Andrew Wallace,Ph.D., Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html ================================================================== From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!www.nntp.primenet.com!nntp.primenet.com!europa.clark.net!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!baron.netcom.net.uk!netcom.net.uk!nntpfeed.doc.ic.ac.uk!sunsite.doc.ic.ac.uk!qmw!alpha.qmw.ac.uk!zgha210 From: Jim Reid Newsgroups: bionet.molbio.proteins Subject: Reversable inhibition of Cys proteases Date: Thu, 5 Jun 1997 12:04:50 +0100 Organization: Queen Mary & Westfield College, London, UK Message-ID: NNTP-Posting-Host: alpha.qmw.ac.uk Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Lines: 27 A good reversible covalent inhibitor of Cysteine proteases is 2PDS (2,2'-dipyridyl disulphide) sold by aldrich as 'Aldrithiol-2' We use this stuff extensively here to block cysteine proteases in purification and storage to prevent autolysis (we also use it and derivatives for kinetic studies but thats a longer story). 2PDS can be made up to about 1.5 mM in water (a saturated soln) if you want a higher conc add some ethanol, If you want to clean up the commercial stuff it can be recrystalysed from ethanol. If you want to regenerate youre thiol group use 20 mM Cys and then G-25 it or something to get rid of the Cys. Have a look at: Brocklehurst. K, Methods in Enzymology (1982) 87 427-469 and Baines, B.S. and Brocklehurst, K. BJ (1978) 173, 345-347 and some purification procedures based on this stuff; Brocklehurst, K. et al Methods in Enzymology (1974) 531-544 [many other good, more recent, reviews on this but this is the only one I can find at the moment] Rich et al BJ (1986) 235, 731-734 [haven't tried this but it looks good] I hope this is of some use, Jim Reid ha4252@qmw.ac.uk From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!agate!hammer.uoregon.edu!vixen.cso.uiuc.edu!chi-news.cic.net!mr.net!newshub.tc.umn.edu!newsstand.tc.umn.edu!limerick.cbs.umn.edu!hiroki From: hiroki@limerick.cbs.umn.edu (Hiroki) Newsgroups: bionet.molbio.proteins Subject: Re: Tissue homogenizer for Eppies? Date: 5 Jun 1997 07:09:18 GMT Organization: University of Minnesota Lines: 22 Distribution: world Message-ID: <5n5omu$r11@epx.cis.umn.edu> References: <199706022122.PAA55078@hector.NMSU.Edu> Reply-To: hiroki@limerick.cbs.umn.edu NNTP-Posting-Host: limerick.cbs.umn.edu X-Newsreader: TIN [version 1.2 PL2] For what it's worth, you can find a similar twaddler for about $5 or so at Target -- a chain store like Walmart, and you need to kludge an adapter to take the little blue pestles. Not sure what it's for, cuticle polisher? nose hair remover? Hiroki Hiranya Roychowdhury (hroychow@NMSU.EDU) wrote: : At 08:48 AM 6/2/97 +0000, Keld Sorensen wrote: : >I think Kontes sell these with a little motorized pestle. : > : >Keld. : > : > : We had gone ahead and bought one of those 'motoized' things. It is useless when you are grinding tissues frozen in liq N2. : Dr. Hiranya Sankar Roychowdhury : Plant Genetic Engineering Lab. : New Mexico State University : Las Cruces, NM 88003 : Ph. (505) 646-5785 : hroychow@nmsu.edu From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!biosci!not-for-mail From: newera@plaza.snu.ac.kr Newsgroups: bionet.molbio.proteins, Subject: Acetic acid + NaOH = buffer at pH 5.0? Date: 5 Jun 1997 09:36:10 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: <5n6ptq$3cl@net.bio.net> NNTP-Posting-Host: net.bio.net Summary: Keywords: A mixture of acetic acid and NaOH is a buffer, especially in the case of a concentrated protein solution around 1~5mM? I am sure that 'Acetic acid + Sodium Acetate = buffer at pH 5.0', but not convinced of 'Acetic acid + NaOH = buffer at pH 5.0' because it is not a mixture of weak acid and its salt form. I am definitely in the situation where the latter is much preferred if it can be a buffer at all. Any comment will be appreciated. Best regards, Lee -- Lee, Ji Hyun newera@plaza.snu.ac.kr Laboratory of Physical Pharmacy(Prof. Lee, Bong Jin) College of Pharmacy Seoul National University Shinlim-Dong, Kwanak-Gu Seoul 151-742, Korea. Tel : 82-2-880-7869 Fax : 82-2-872-3632 From owner-proteins@net.bio.net Wed Jun 04 23:00:00 1997 Path: biosci!agate!howland.erols.net!newsfeed.internetmci.com!in3.uu.net!138.133.17.7!amgen!usenet From: John Philo Newsgroups: bionet.molbio.proteins, Subject: Re: Acetic acid + NaOH = buffer at pH 5.0? Date: Thu, 05 Jun 1997 13:20:20 -0700 Organization: Amgen Inc. Lines: 33 Message-ID: <33971F84.4F02@amgen.com> References: <5n6ps9$2no@net.bio.net> Reply-To: jphilo@amgen.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01Gold (Win95; I) To: newera@plaza.snu.ac.kr newera@plaza.snu.ac.kr wrote: > > Is a mixture of acetic acid and NaOH a buffer, especially in a concentrated protein solution around 1~5mM? > > I am sure that 'Acetic acid + Sodium Acetate = buffer at pH 5.0', > but not convinced of 'Acetic acid + NaOH = buffer at pH 5.0' because it is not a mixture of weak acid and its salt form. > > I am definitely in the situation where the latter is much preferred if it can be a buffer at all. > > Any comment will be appreciated. > > Best regards, > > Lee, Ji Hyun newera@plaza.snu.ac.kr > Laboratory of Physical Pharmacy(Prof. Lee, Bong Jin) > College of Pharmacy > Seoul National University You are mistaken. A mixture of 'acetic acid + sodium acetate' is entirely equivalent to that of 'acetic acid + NaOH', at the same pH and total concentration of acetate ion. You will end up with the same concentrations of ions no matter which way you get there. Therefore the latter cannot be "preferred" over the former, unless you simply don't have sodium acetate on hand! You don't say what concentration of acetate you intend to use, but at low buffer concentrations (say 10 millimolar) it is likely that your 1-5 mM protein will dominate the overall buffering capacity of the solution, since the concentration of carboxylate groups and histidine residues will probably greatly exceed that of buffer. John Philo, Protein Chemistry, Amgen *** Disclaimer: These are the opinions of the poster not Amgen Inc.*** From owner-proteins@net.bio.net Thu Jun 05 23:00:00 1997 Path: biosci!GUARANY.CPD.UNB.BR!zanotta From: zanotta@GUARANY.CPD.UNB.BR (pedro jose portugal zanotta) Newsgroups: bionet.molbio.proteins Subject: Re: Acetic acid + NaOH = buffer at pH 5.0? Date: 6 Jun 1997 04:53:49 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <5n6ps9$2no@net.bio.net> NNTP-Posting-Host: net.bio.net The key point is that you have, in this pH, a misture of acetic acid plus acetate (which is the conjugate base of th acid). So, you would have a buffer. Hope this helps. Pedro Zanotta From owner-proteins@net.bio.net Thu Jun 05 23:00:00 1997 Path: biosci!EXMDI1.MPIEM.GWDG.DE!hesse From: hesse@EXMDI1.MPIEM.GWDG.DE ("D. Hesse") Newsgroups: bionet.molbio.proteins Subject: Problems with PTH-Columns supplied by PE/ABI Date: 6 Jun 1997 06:40:51 -0700 Organization: MPI f|r experimentelle Medizin Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: <339813A4.121D@exmdi1.mpiem.gwdg.de> NNTP-Posting-Host: net.bio.net Here is a question about the PTH-Columns supplied by Perkin-Elmer/Applied Biosystems : In former times we had a lifetime of nearly 1500 ( and more ) injections per column. Then, column lifetime decreased rapidly to 500 - 700 injections per column. In the same period , we noticed the first time a white material flushing from the column when it was equilibrated . This white material was insoluble in different solvents. Fortunately, we wash every column first without a connection to the uv-flowcell. Now,this material has another property, it is soluble ( in Solvent A3 + Premix and Solvent B2 ), and may pass the detector flow cell without plugging . But the columns require in our hands larger amounts of Premix concentrate to fix the positions of Histidine and Arginine. Has anyone else also observed this behaviour of the columns in his lab? Thanks for help. -- Doerte Hesse Max-Planck-Institut f|r experimentelle Medizin Abt. Immunchemie Hermann-Rein-Strasse 3 37075 Goettingen Tel.: 0551 / 3899-304 Fax : 0551 / 3899-323 E-mail : Hesse@exmdi1.mpiem.gwdg.de From owner-proteins@net.bio.net Thu Jun 05 23:00:00 1997 Path: biosci!agate!hammer.uoregon.edu!csulb.edu!info.ucla.edu!nnrp.info.ucla.edu!usenet From: Edmundo Castro Newsgroups: bionet.molbio.proteins Subject: Commercial peptide synthesis Date: Thu, 05 Jun 1997 09:56:49 -0700 Organization: UCLA Lines: 6 Message-ID: <3396EFD0.1185@physci.ucla.edu> Reply-To: ecastro@physci.ucla.edu NNTP-Posting-Host: 149.142.241.23 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.01 (Macintosh; I; PPC) Can someone recommend a company that synthesizes peptides? We are interested in obtaining one gram of a decapeptide, no modifications. Thanks in advance, E. Castro-Vargas From owner-proteins@net.bio.net Thu Jun 05 23:00:00 1997 Path: biosci!rutgers.rutgers.edu!gatech!csulb.edu!hammer.uoregon.edu!news-xfer.netaxs.com!news.maxwell.syr.edu!howland.erols.net!vixen.cso.uiuc.edu!uwm.edu!newsspool.doit.wisc.edu!news.doit.wisc.edu!alpha.oncology.wisc.edu!user From: mcmahan@oncology.wisc.edu (Scott McMahan) Newsgroups: bionet.molbio.proteins, Subject: Re: Acetic acid + NaOH = buffer at pH 5.0? Date: Fri, 06 Jun 1997 00:20:44 -0500 Organization: University of Wisconsin-Madison Lines: 22 Distribution: world Message-ID: References: <5n6ptq$3cl@net.bio.net> NNTP-Posting-Host: alpha.oncology.wisc.edu X-Newsreader: Yet Another NewsWatcher 2.1.8 In article , pxpst2+@pitt.edu (peter) wrote: :In article <5n6ptq$3cl@net.bio.net>, newera@plaza.snu.ac.kr wrote: : :.> Summary: :.> Keywords: :.> :.> A mixture of acetic acid and NaOH is a buffer, especially in the case :of a .concentrated protein solution around 1~5mM? : :Protein in solution has nothing to do with whether or not that is a :buffer. As for your question in is a buffer system but not a good one :because you are using a strong base with a strong acid At pH 5.0 (as mentioned in the subject header), which is close to the pKa (4.8 I think) of acetate, there should be a significant amount of both the acid and base forms of the acetate, giving good buffering capacity. -- Scott McMahan mcmahan@oncology.wisc.edu From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!cam-news-hub1.bbnplanet.com!news.bbnplanet.com!news-feed1.tiac.net!uunet!uunet!194.119.128.129!news.u-net.com!not-for-mail From: awilson@aw.u-net.com (A Wilson) Newsgroups: bionet.molbio.proteins Subject: VOTING IN PROGRESS for creation of new newsgroup sci.bio.immunocytochem Date: Mon, 09 Jun 1997 20:21:46 GMT Organization: U-NET Ltd Lines: 41 Message-ID: <5nhola$l20$11@despair.u-net.com> Reply-To: awilson@aw.u-net.com NNTP-Posting-Host: p38.ascend3.is2.u-net.net X-Newsreader: Forte Free Agent 1.0.82 VOTING IN PROGRESS FOR IMMUNOCYTOCHEMISTRY NEWSGROUP sci.bio.immunocytochem If you want to see a newsgroup dedicated to immunohistochemistry, immunocytochemistry and other affinity labelling methods, then please do try to vote if you havn't already done so. The votetaker must collect 100 more YES than NO votes for the group to be made official. The closing date for ballots to reach the votetaker is 16th June. All you need to vote is a valid e-mail address. The 2nd (and final) CFV, or CALL FOR VOTES was posted to "news.announce.newgroups" and "news.groups" on the 6th June. Voting instructions and the ballot can be found at the end of the CFV. Some people are having trouble accessing the CFV or "Call For Votes" via the Usenet Newsgroups. The VOTETAKER, David Bostwick, will SEND YOU the CFV (with voting instructions and the ballot) if you e-mail him bostwick@cas.chemistry.gatech.edu Alternatively, you can go the the FTP site: ftp://ftp.isc.org/pub/usenet/news.announce.newgroups/sci/sci.bio.immunocytochem where all the old "Request For Discussions" are stored, together with the CFV (right at the end!) Remember, the closing date for votes to reach the votetaker is 16th June, so hurry! Please feel free to e-mail me with any questions you may have about the proposed group or voting. Amanda Wilson awilson@aw.u-net.com Deputy Manager, Electron Microscope Unit, St George's Hospital Medical School London UK Tel: 0181 725 5220 e-mail From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!delos.dra.hmg.gb!server1.netnews.ja.net!lyra.csx.cam.ac.uk!powercenter-604.physiol.cam.ac.uk!user From: whc23@cus.cam.ac.uk (Dr.W.H. Colledge) Newsgroups: bionet.molbio.proteins Subject: autohaemolysis assay Date: Mon, 09 Jun 1997 09:28:34 +0100 Organization: University of Cambridge Message-ID: NNTP-Posting-Host: powercenter-604.physiol.cam.ac.uk Lines: 12 Does anyone have a method or can give me a reference for measuring the autohaemolysis of red blood cells? Thanks. -- Dr. W.H. Colledge Physiology University of Cambridge Cambridge CB2 3EG Tel. 01223-333881 Fax. 01223-333840 From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!howland.erols.net!wits.uni.net.za!ru.uni.net.za!und.uni.net.za!und.ac.za!cc.unp.ac.za!nobody From: nobody@cc.unp.ac.za Newsgroups: bionet.molbio.proteins Subject: anti-IgE Date: Mon, 9 Jun 1997 14:02:59 GMT Organization: University of Natal , Pietermaritzburg, South Africa Lines: 10 Message-ID: NNTP-Posting-Host: unpsun3.cc.unp.ac.za Cache-Post-Path: unpsun3.cc.unp.ac.za!unknown@143.128.128.139 Hi all, Does anybody have any anti-rabbit-IgE or anti-rat-IgE that they may be willing to part with? I would be most grateful if you would contact me. Many thanks in advance, Rory Morty e-mail: MortyR@biochem.unp.ac.za From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!news.bbnplanet.com!newsfeed.nacamar.de!nntp.uio.no!news2.interlog.com!news.interlog.com!yblee From: samuel.lee@utoronto.ca (Samuel P. Lee) Newsgroups: bionet.molbio.proteins Subject: Re: SDS electroelution Date: Sat, 07 Jun 97 17:26:01 GMT Organization: University of Toronto Lines: 22 Message-ID: <5nc5sd$3hg@news.interlog.com> References: <5n9lmq$cb7@news.bu.edu> NNTP-Posting-Host: ip213-231.cc.interlog.com X-Newsreader: News Xpress Version 1.0 Beta #4 In article <5n9lmq$cb7@news.bu.edu>, akopian@bu.edu wrote: >Can someone send me protocol for removal (elution) of proteins from >SDS-PAGE without having to by the electroelution equipment I've heard >so much about. Is there a quick and simple technique for removing >proteins from PAGE gels? Thanks in advance...please respond to >akopian@bu.edu. In our lab, we have successfully eluted proteins from PAGE gels by doing the following: Cut out the area of gel that contains the protein of interest. Place the gel slice in an micro test tube. Add several volumes of an appropriate buffer (one that contains SDS works best). Crush or grind up the gel and allow the mixture to sit for 1 to 4 hours. Centrifuge and remove the supernatant which should contain a significant amount of protein. We have found this simple diffusion method works quite well. Sam Lee Department of Pharmacology University of Toronto samuel.lee@utoronto.ca From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Newsgroups: bionet.molbio.proteins Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!delos.dra.hmg.gb!server1.netnews.ja.net!server5.netnews.ja.net!server6.netnews.ja.net!str-ccsun!strath-cs!liv!news From: lgbell@liverpool.ac.uk Subject: Cardiac Myosins. Content-Type: text/plain; charset=us-ascii Message-ID: <33980F55.69AC@liverpool.ac.uk> Sender: news@liverpool.ac.uk (News System) Nntp-Posting-Host: pc028010.med.liv.ac.uk Content-Transfer-Encoding: 7bit Organization: The University of Liverpool Mime-Version: 1.0 Date: Fri, 6 Jun 1997 13:23:33 GMT X-Mailer: Mozilla 2.02 (Win16; I) Lines: 10 Wanted structural information on mammalian cardiac myosins. 3D and crystal structure data is apparrently sparse both at Brrohaven and SWISS-PROT, The nearest being chicken smooth muscle I believe. Any extra information would be wellcome. Thanks in advance for your assistance. Len Bell lgbell@liv.ac.uk From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!europa.clark.net!newsfeeds.sol.net!portc02.blue.aol.com!newsfeed.pitt.edu!pelli.pathology.pitt.edu!user From: pxpst2+@pitt.edu.delete (peter) Newsgroups: bionet.molbio.proteins, Subject: Re: Acetic acid + NaOH = buffer at pH 5.0? Date: 6 Jun 1997 19:38:17 GMT Organization: Univ.of Pittsburgh Lines: 20 Distribution: world Message-ID: References: <5n6ptq$3cl@net.bio.net> NNTP-Posting-Host: pelli.pathology.pitt.edu .In article , .mcmahan@oncology.wisc.edu (Scott McMahan) wrote: .> In article , .> pxpst2+@pitt.edu (peter) wrote: .> .> :In article <5n6ptq$3cl@net.bio.net>, newera@plaza.snu.ac.kr wrote: .> : .> :.> Summary: .> :.> Keywords: .> : .> :Protein in solution has nothing to do with whether or not that is a .> :buffer. As for your question in is a buffer system but not a good one .> :because you are using a strong base with a strong acid . > I mistook acetic acid for a strong acid but it does not ionize completely therefore it is not a strong acid sorry for the mis information Peter From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cam-news-feed2.bbnplanet.com!news.bbnplanet.com!news.bu.edu!usenet From: akopian@bu.edu Newsgroups: bionet.molbio.proteins Subject: SDS electroelution Date: 6 Jun 1997 18:42:34 GMT Organization: bu Lines: 5 Message-ID: <5n9lmq$cb7@news.bu.edu> Reply-To: akopian@bu.edu NNTP-Posting-Host: med-arth53.bu.edu X-Newsreader: WinVN 0.92.6+ Can someone send me protocol for removal (elution) of proteins from SDS-PAGE without having to by the electroelution equipment I've heard so much about. Is there a quick and simple technique for removing proteins from PAGE gels? Thanks in advance...please respond to akopian@bu.edu. From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!agate!hammer.uoregon.edu!news-xfer.netaxs.com!europa.clark.net!newsxfer3.itd.umich.edu!newsxfer.itd.umich.edu!news.mtu.edu!msunews!harbinger.cc.monash.edu.au!microb36.med.monash.edu.au!CAUCHI From: CAUCHI@med.monash.edu.au (Mark Cauchi) Newsgroups: bionet.molbio.proteins Subject: Recombinant DNA Techniques Course Date: Mon, 9 Jun 1997 06:56:52 GMT Organization: Monash University Lines: 8 Distribution: AU Message-ID: NNTP-Posting-Host: microb36.med.monash.edu.au X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] The Micromon Unit at the Department of Microbiology, Monash University in Melbourne, Australia will be running its Recombinant DNA Techniques Course between the 16-21 November, 1997. This is an introductory-intermediate level course which offers a skills-based training package. If you would like further information, details can be found on our web page at http://www.monash.edu.au/informatics/micro/department/dnacorse.htm From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!newsfeed.internetmci.com!jump.net!grunt.dejanews.com!not-for-mail Date: Sun, 08 Jun 1997 03:11:12 -0600 From: nvl@chez.com Subject: wheat glutathione reductase Newsgroups: bionet.molbio.proteins Message-ID: <865757417.10600@dejanews.com> Reply-To: nvl@chez.com Organization: Deja News Usenet Posting Service X-Article-Creation-Date: Sun Jun 08 08:10:17 1997 GMT X-Originating-IP-Addr: 195.154.5.178 (ppp39.mon.hol.fr) X-Http-User-Agent: Mozilla/2.02E [fr]-HAVAS1 (Win95; I) X-Authenticated-Sender: nvl@chez.com Lines: 8 as anybody heard about wheat glutathione reductase ? I would be very interested in. Thank you Nicolas Vianey-Liaud -------------------==== Posted via Deja News ====----------------------- http://www.dejanews.com/ Search, Read, Post to Usenet From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!cam-news-hub1.bbnplanet.com!news.bbnplanet.com!news-feed1.tiac.net!uunet!uunet!194.119.128.129!news.u-net.com!not-for-mail From: awilson@aw.u-net.com (A Wilson) Newsgroups: bionet.molbio.proteins Subject: VOTING IN PROGRESS for sci.bio.immunocytochem Date: Mon, 09 Jun 1997 20:21:32 GMT Organization: U-NET Ltd Lines: 30 Message-ID: <5nhoks$l20$2@despair.u-net.com> Reply-To: awilson@aw.u-net.com NNTP-Posting-Host: p38.ascend3.is2.u-net.net X-Newsreader: Forte Free Agent 1.0.82 ATTENTION ALL IMMUNOHISTOCHEMISTS, IMMUNOCYTOCHEMISTS AND OTHER AFFINITY LABELLERS!!! VOTING IS NOW IN PROGRESS FOR A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP! The official CFV or CALL FOR VOTES for the proposed new newsgroup has recently been posted to news.announce.newgroups and news.groups. If you are keen to see a newsgroup dedicated to discussion of immunocytochemistry and related topics, then please do vote. All you need to qualify as a voter is a valid e-mail address. To vote for the new group, find the official posting named (posted on 26th May 1997) in news.announce.newgroups or news.groups, and look for the VOTING INSTRUCTIONS and BALLOT at the end of the CFV. You MUST use the official ballot to vote, and it MUST be posted to the official votetaker for it to be counted. Thank-you. Amanda Wilson, e-mail Proponent of the proposed newsgroup Deputy Manager, Electron Microscope Unit, St George's Hospital Medical School London UK Tel: 0181 725 5220 e-mail From owner-proteins@net.bio.net Sun Jun 08 23:00:00 1997 Path: biosci!news.Stanford.EDU!agate!hammer.uoregon.edu!xfer.kren.nm.kr!nntp.kreonet.re.kr!newsfeed.dacom.co.kr!newsfeed.internetmci.com!feeder.chicago.cic.net!newsxfer.nether.net!141.211.144.12!newsxfer.itd.umich.edu!uunet!uunet!194.119.128.129!news.u-net.com!not-for-mail From: awilson@aw.u-net.com (A Wilson) Newsgroups: bionet.molbio.proteins Subject: VOTING IN PROGRESS for creation of new newsgroup sci.bio.immunocytochem Date: Mon, 09 Jun 1997 19:41:04 GMT Organization: U-NET Ltd Lines: 41 Message-ID: <5nhm91$jbt$2@despair.u-net.com> Reply-To: awilson@aw.u-net.com NNTP-Posting-Host: p16.ascend4.is2.u-net.net X-Newsreader: Forte Free Agent 1.0.82 VOTING IN PROGRESS FOR IMMUNOCYTOCHEMISTRY NEWSGROUP sci.bio.immunocytochem If you want to see a newsgroup dedicated to immunohistochemistry, immunocytochemistry and other affinity labelling methods, then please do try to vote if you havn't already done so. The votetaker must collect 100 more YES than NO votes for the group to be made official. The closing date for ballots to reach the votetaker is 16th June. All you need to vote is a valid e-mail address. The 2nd (and final) CFV, or CALL FOR VOTES was posted to "news.announce.newgroups" and "news.groups" on the 6th June. Voting instructions and the ballot can be found at the end of the CFV. Some people are having trouble accessing the CFV or "Call For Votes" via the Usenet Newsgroups. The VOTETAKER, David Bostwick, will SEND YOU the CFV (with voting instructions and the ballot) if you e-mail him bostwick@cas.chemistry.gatech.edu Alternatively, you can go the the FTP site: ftp://ftp.isc.org/pub/usenet/news.announce.newgroups/sci/sci.bio.immunocytochem where all the old "Request For Discussions" are stored, together with the CFV (right at the end!) Remember, the closing date for votes to reach the votetaker is 16th June, so hurry! Please feel free to e-mail me with any questions you may have about the proposed group or voting. Amanda Wilson awilson@aw.u-net.com Deputy Manager, Electron Microscope Unit, St George's Hospital Medical School London UK Tel: 0181 725 5220 e-mail From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 Path: biosci!news.Stanford.EDU!agate!hammer.uoregon.edu!newsfeed.internetmci.com!europa.clark.net!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!rill.news.pipex.net!pipex!server1.netnews.ja.net!server5.netnews.ja.net!daresbury!usenet From: jamie bickley Newsgroups: bionet.molbio.proteins Subject: aa analysis Date: Tue, 10 Jun 1997 09:17:04 +0100 Organization: clrc Distribution: bionet Message-ID: <339D0D80.7605@dl.ac.uk> References: <338F4323.3993@bris.ac.uk> Reply-To: jfb@dl.ac.uk NNTP-Posting-Host: jhavig.dl.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0Gold (Win16; I) Lines: 2 hi, could anyone tell me what the size limit of polypeptide is for amino acid analysis on hplc?? cheers From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 Path: biosci!rutgers!gatech!csulb.edu!hammer.uoregon.edu!newsfeed.internetmci.com!howland.erols.net!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!delos.dra.hmg.gb!server1.netnews.ja.net!warwick!bham!not-for-mail From: altabios@bham.ac.uk (John E. Fox) Newsgroups: bionet.molbio.proteins Subject: Re: Q: BrCN-cleavage in a gel? Date: 10 Jun 1997 11:57:02 GMT Organization: Alta Bioscience Message-ID: <5njfee$qqt$1@usenet.bham.ac.uk> References: NNTP-Posting-Host: bcs118.bham.ac.uk X-Newsreader: WinVN 0.90.6 Lines: 16 In article , r103@bham.ac.uk (room 103) says: > >Hi there, > >I am interested in a method to cleave a protein with a molecular mass >300 >kDa with BrCN. Enzymatic digests would lead to more than 400 fragments, >therefore I think BrCN might be the best choice. Although I could blot and Give me a call Peter. I have a method. John Fox ****************************************************************** Alta BioScience Email: altabios@bham.ac.uk School of Biochemistry Phone: 0121-414-5450 The University of Birmingham Fax: 0121-414-3376 Edgbaston, BIRMINGHAM, B15 2TT, UK From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 Path: biosci!UMBI.UMD.EDU!collins From: collins@UMBI.UMD.EDU (John Collins) Newsgroups: bionet.molbio.proteins Subject: Re: -Hot Teens that want to be... Date: 10 Jun 1997 22:07:38 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 9 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <5njdak$5vv$3@news9.gte.net> NNTP-Posting-Host: net.bio.net On 10 Jun 1997 afdl@umbi.umd.edu wrote: > Check out this site, it has tons of Nude Teenagers... ==================================================================== Attention all subscribers! This offensive message did not come from anyone at umbi. The "from" address was faked. Is there no way to stop this sort of thing? -John Collins From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 Path: biosci!UMBI.UMD.EDU!collins From: collins@UMBI.UMD.EDU (John Collins) Newsgroups: bionet.molbio.proteins Subject: Re: CABLE BOX DESCRAMBLER BUILD YOUR OWN DESCRAMBLER FAST CHEAP AND EASY Date: 10 Jun 1997 22:07:10 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <5nknc6$e72@chronicle.concentric.net> NNTP-Posting-Host: net.bio.net This group needs a monitor to filter out this sort of trash. On 10 Jun 1997, Free Cable wrote: > CABLE BOX DESCRAMBLER BUILD YOUR OWN DESCRAMBLER FAST CHEAP AND EASY > > > Just a few inexpensive parts from Radio Shack and a little time and you can descramble every cable channel. See all your favorite movie channels,pay per view etc > > To recieve detail instructions and diagrams on how to construct your own Cable Box Descrambler > Mail $5.00 CASH MONEY ORDER CHECK > > S&G Enterprise > 12145 Augusta Woods Cir > Suite 3 > Orlando FL. 32824 > > Please be sure to include your full name & address > Allow 10 days to recieve > > Thank You > Seth Garner > > > > > From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 Path: biosci!MSU.EDU!RNREUSCH From: RNREUSCH@MSU.EDU ("Rosetta N. Reusch") Newsgroups: bionet.molbio.proteins Subject: Poly(3-hydroxybutyrate) modification of proteins Date: 10 Jun 1997 21:58:21 -0700 Organization: MSU Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <339D90D8.5DA3F27C@msu.edu> Reply-To: RNREUSCH@msu, edu@server.mph.msu.edu NNTP-Posting-Host: net.bio.net We are finding that a number of proteins are modified by short-chain poly(3-hydroxybutyrate) (cPHB). cPHB is invisible in the UV, cannot be observed by X-ray crystallography, and is removed by conditions used in sequencing. If you want to know whether your protein is a cPHB conjugate and can spare a few ug of pure protein, contact me. We have antibody to synthetic PHB and can determine this quickly by Western blot analysis. Rosetta Reusch RNREUSCH@msu.edu From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 From: lhom@nature.berkeley.edu (Louis Hom) Newsgroups: bionet.cellbiol,bionet.molbio.proteins Subject: N-linked glycosylation Date: 10 Jun 1997 20:43:04 GMT Organization: University of California, Berkeley Lines: 9 Message-ID: <5nke8o$b3p@agate.berkeley.edu> NNTP-Posting-Host: nature.berkeley.edu Path: biosci!rutgers!gatech!news1.mid-ga.com!news.hom.net!nntp.mid-ga.com!news.oru.edu!news-spur1.maxwell.syr.edu!news.maxwell.syr.edu!newsfeed.kornet.nm.kr!agate!nature.berkeley.edu!lhom Xref: biosci bionet.cellbiol:7475 bionet.molbio.proteins:10941 In Alberts, they say that the initial attachment of the 14-sugar complex to Asn's in the ER happens rapidly . . . something like "most proteins are glycosylated as they are translocated". Anyway, the important part was the "most" -- does anyone have an idea how much "most" is? -- _______________________________________________________________________________ Lou Hom >K '93 "No one ever went broke lhom@nature.berkeley.edu underestimating the taste http://www.ocf.berkeley.edu/~lhom of the American public." --Mencken. From owner-proteins@net.bio.net Mon Jun 09 23:00:00 1997 Path: biosci!rutgers!gatech!csulb.edu!hammer.uoregon.edu!newsfeed.internetmci.com!howland.erols.net!psinntp!news.columbia.edu!aloha.cc.columbia.edu!mdt1 From: M David Tilson Newsgroups: bionet.molbio.proteins Subject: re: Need help with ProAnWin Date: Tue, 10 Jun 1997 15:07:39 -0400 Organization: Columbia University Lines: 27 Message-ID: NNTP-Posting-Host: aloha.cc.columbia.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII I'm tired of waiting around for pairwise and multiple amino acid sequence alignments to run on the big servers, so I've downloaded ProAnWin to try to do alignments on my PC. The "Getting Started" section of Help doesn't give me enough info to get going. Under "File" you can request loading of a .seq file; but how do you get your sequences of interest into that first .seq file. I can't find any cut and paste functions to help get started. Anybody know of other software that is more intuitive. Thanks in advance, _dave_ E-mail: mdt1@columbia.edu URL : http://www.columbia.edu/~mdt1/ 1 = one (not little L), and don't forget the trailing "/") Phone : 212-523-7779 SnailM: M David Tilson, SLRHC, 1000 Tenth Avenue, NY, NY 10019 From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!rutgers!gatech!howland.erols.net!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!delos.dra.hmg.gb!server1.netnews.ja.net!server5.netnews.ja.net!daresbury!is.bbsrc.ac.uk!news From: Andy Phillips Newsgroups: bionet.molbio.proteins Subject: Re: Need help with ProAnWin Date: Wed, 11 Jun 1997 09:10:54 +0100 Organization: BBSRC, IACR Long Ashton Research Station Message-ID: <339E5D8E.7B7970F4@bbsrc.ac.uk> References: Reply-To: andy.phillips@bbsrc.ac.uk NNTP-Posting-Host: pc0628.lars.bbsrc.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.0b5 [en] (Win95; I) CC: andy.phillips@bbsrc.ac.uk X-Priority: 3 (Normal) Lines: 35 M David Tilson wrote: > > I'm tired of waiting around for pairwise and multiple amino acid > sequence alignments to run on the big servers, so I've downloaded > ProAnWin to try to do alignments on my PC. The "Getting Started" > section of Help doesn't give me enough info to get going. Under "File" > you can request loading of a .seq file; but how do you get your > sequences of interest into that first .seq file. I can't find any > cut and paste functions to help get started. > > Anybody know of other software that is more intuitive. > > Thanks in advance, > > _dave_ > > E-mail: mdt1@columbia.edu I use ClustalX - see the link on my molbiol web page (http://www.lars.bbsrc.ac.uk/plantsci/molbiol/molbiol.html) under the software section. This runs under Win95 and is great for generating multiple alignments. I haven't tried it for pairwise alignments, but I see no reason why it shouldn't work. For manual tidying-up and shaded output I use Genedoc (again there's a link on my web page) Andy ------------------------------------------------------------------------ Email : andy.phillips@bbsrc.ac.uk : University of Bristol Home : andy@cycad.demon.co.uk : IACR Long Ashton Research Station Phone : +44-1275-549257 : Long Ashton Fax : +44-1275-394281 : Bristol, BS18 9AF, UK WWW : http://www.lars.bbsrc.ac.uk/plantsci/molbiol/molbiol.html ------------------------------------------------------------------------ From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!BIO.VU.NL!ahdeboer From: ahdeboer@BIO.VU.NL ("A.H. de Boer") Newsgroups: bionet.molbio.proteins Subject: Postdoc position (14-3-3 proteins) open in Amsterdam Date: 11 Jun 1997 15:51:53 -0700 Organization: Vrije Universiteit Amsterdam Lines: 44 Sender: daemon@net.bio.net Distribution: world Message-ID: <339FAB7E.15A3@bio.vu.nl> NNTP-Posting-Host: net.bio.net A postdoctoral position is available to investigate the role of 14-3-3 proteins in the regulation of the plant plasma membrane H+-ATPase and nitrate reductase, an enzyme which is located in the cytosol. 14-3-3 proteins play a key role in coordinating multiple signalling pathways (see Aitken 1996, Trends Cell Biol. 6, 341-347) by serving as docking proteins for receptors, kinases and phosphatases. In plants some 14-3-3 isoforms make up the receptor for the phytotoxin fusicoccin (see de Boer 1997, Trends Plant Sci. 2, 60-66). Plants have a 14-3-3 gene family with 6 to 10 members and it has been speculated that one isoform has more than one function. Using a combination of biochemical, genetic and molecular approaches we want to address the question which isoforms bind to (and control the activity of) the ATPase and nitrate reductase respectively. Since activation of root nitrate reductase also activates the ATPase, 14-3-3 dimers may mediate the ‘cross-talk' between these two enzymes. The dynamic behaviour of 14-3-3 isoforms (viz. translocation within the cell and docking to the respective targets proteins) will be studied by means of Green Fuorescent Protein (GFP) constructs. This project, is part of an EU Framework IV Biotech-programme entitled ‘Central Role in Adaptation of Fourteen Three Three proteins', and is carried out by six international partners. In the lab you will be part of a group of 3 postdocs working on 14-3-3 proteins in barley and Petunia. Candidates should have demonstrated experience and publications in biochemistry/protein chemistry and molecular biology. Experience with microscopic techniques is considered favourably. The position will be for a period of 3 years in the Department of Genetics, Section Plant Physiology of The Vrije Universiteit Amsterdam, The Netherlands. Applicants should send (before July 10) a curriculum vitae, a brief statement of research interest and names and phone numbers and e-mail address of 3 references to: Bert de Boer Faculty of Biology, Vrije Universiteit Amsterdam Dep. Of Genetics, Section Plant Physiology De Boelelaan 1087 1081 HV Amsterdam, the Netherlands. Phone: 31-20- 444 7162 Fax: 31-20-444 7229 E-mail: ahdeboer@bio.vu.nl PS: I am out of the country until July 5th, but after that I will be happy to give more information about the position. From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!bmg.bhs.uab.edu!GMEACHAM From: GMEACHAM@bmg.bhs.uab.edu Newsgroups: bionet.molbio.proteins Subject: Looking for a commercial supplier of SRP Date: 11 Jun 1997 08:51:38 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: <143696C5603@bmg.bhs.uab.edu> NNTP-Posting-Host: net.bio.net Does anyone know of a supplier of purified SRP (signal recognition particle) or the cDNA for SRP? Any information would be helpful. Thanks in advance. Geoff Meacham UAB Dept. of Cell Biology (205) 975-4893 From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!rutgers!gatech!csulb.edu!hammer.uoregon.edu!newsfeed.direct.ca!news-peer.gsl.net!news-hk.gsl.net!news.gsl.net!newsgate.cuhk.edu.hk!news.hku.hk!news From: Alan Lee Newsgroups: bionet.molbio.proteins Subject: taurine assay Date: Wed, 11 Jun 1997 19:10:51 +0800 Organization: Institute of Molecular Biology, University of Hong Kong Lines: 8 Message-ID: <339E87BB.7E58@hkucc.hku.hk> Reply-To: ywlee@hkucc.hku.hk NNTP-Posting-Host: 147.8.72.23 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (Win95; I) Hello, I am currently looking for protocols for HPLC determination of taurine in biological samples. The ideal protocol would be able to exclude other amino acids except taurine. Anyone came across such protocols please kindly let me know. Thamk you very much. Alan Lee. e-mail: ywlee@hkucc.hku.hk June 11, 97 From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!agate!howland.erols.net!europa.clark.net!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!delos.dra.hmg.gb!server1.netnews.ja.net!news.qub.ac.uk!not-for-mail From: Andrew Wallace Newsgroups: bionet.molbio.proteins Subject: Re: Very Long Polypeptides Date: Wed, 11 Jun 1997 11:24:05 +0100 Organization: Queens University Belfast Message-ID: <339E7CC3.5B75@qub.ac.uk.see.signature> References: <5nlsi7$ggv@lyra.csx.cam.ac.uk> Reply-To: a.wallace@qub.ac.uk.see.signature NNTP-Posting-Host: awall.bc.qub.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (Macintosh; I; PPC) Lines: 41 P.A. Watters wrote: > > Hi, > > I am interested in looking at some examples of the primary > structure of very long polypeptides to test some numerical > methods with. I'd appreciate any examples that could be > forwarded! > > ASCII or RASMOL format would be fine. > > Best Wishes, > Paul > > -- > Paul A. Watters > Physiological Laboratory, University of Cambridge Could you be more specific about what you mean by very long polypeptides, i.e. how many residues are we talking about? I read somewhere that the sequence of the muscle protein titin was completed recently. This protein has a MW of around 1,000,000. Would that be large enough for you? Another large protein you might consider is Von Willebrand Factor, I believe the sequences of the protein from several different species are now in the database. A good place to look for protein sequences and Medline references is the Entrez browser on the web: http://www3.ncbi.nlm.nih.gov/Entrez/ Andrew -- - note antispam feature in return address. My real address is: ================================================================== Andrew Wallace,Ph.D., Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html ================================================================== From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!agate!newsfeed.kornet.nm.kr!xfer.kren.nm.kr!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!disgorge.news.demon.net!demon!dispatch.news.demon.net!demon!delos.dra.hmg.gb!server1.netnews.ja.net!lyra.csx.cam.ac.uk!paw24 From: paw24@cus.cam.ac.uk (P.A. Watters) Newsgroups: bionet.molbio.proteins Subject: Very Long Polypeptides Date: 11 Jun 1997 09:53:11 GMT Organization: University of Cambridge, England Message-ID: <5nlsi7$ggv@lyra.csx.cam.ac.uk> NNTP-Posting-Host: taurus.cus.cam.ac.uk X-Newsreader: TIN [version 1.2 PL2] Lines: 15 Hi, I am interested in looking at some examples of the primary structure of very long polypeptides to test some numerical methods with. I'd appreciate any examples that could be forwarded! ASCII or RASMOL format would be fine. Best Wishes, Paul -- Paul A. Watters Physiological Laboratory, University of Cambridge From owner-proteins@net.bio.net Tue Jun 10 23:00:00 1997 Path: biosci!LISTSERV.STARWAVE.COM!espn-sportszone From: espn-sportszone@LISTSERV.STARWAVE.COM (ESPN-SPORTSZONE) Newsgroups: bionet.molbio.proteins Subject: New! ABCNEWS.com and Enhanced Baseball Coverage! Date: 11 Jun 1997 18:03:08 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 49 Sender: daemon@net.bio.net Distribution: world Message-ID: <01BC7688.60EDD380@ENOS> Reply-To: ESPN SPORTZONE USERS NNTP-Posting-Host: net.bio.net Just a note to let you know about our new enhanced baseball coverage and an incredible new service from Starwave, ABCNEWS.com! ABCNEWS.com: Now Always On Did you ever wish you could get ALL your news from SportsZone? Well, now you can get the same kind of round-the-clock, up-to-the-minute coverage from ABCNEWS.com. This new service from ABC News and Starwave offers an incredible range of world, U.S., and local news. You also get business and financial reports, science and technology headlines, weather, entertainment, and plenty of down-to-earth stuff you need to know just to get by these days. You can even get the latest SportsZone headlines! Check out the big picture at ABCNEWS.com: http://www.abcnews.com SportsZone Baseball: The Ultimate Baseball Resource on the Web Heads up: The big leagues just got bigger. You already know that SportsZone brings you great baseball coverage online. Now you're going to be even happier because we've beefed up your daily scores and stats. Starting soon, you'll be able to follow every Major League game pitch-by-pitch in real time with ScoreTracker, our customizable Java scoreboard. Other great new features include: - Complete Game Day Insider previews for every Major League game; - Complete Game Logs for every player; - Exclusive Daily Performance Charts that let you instantly analyze and compare player performance trends. Step up to SportsZone Baseball: http://espn.sportszone.com/mlb/index.html Thanks for making ESPN SportsZone your online sports service! The SportsZone Team. From owner-proteins@net.bio.net Wed Jun 11 23:00:00 1997 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.erols.net!newsfeed.nacamar.de!news-kar1.dfn.de!news-stu1.dfn.de!news-mue1.dfn.de!news-nue1.dfn.de!uni-erlangen.de!winx03!wpxx02!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins Subject: Re: -Hot Teens that want to be... Date: Wed, 11 Jun 1997 14:30:24 +0200 Organization: University of Wuerzburg, Germany Lines: 54 Distribution: world Message-ID: <0p5mn5.jho.ln@wpxx02.toxi.uni-wuerzburg.de> References: <5njdak$5vv$3@news9.gte.net> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] John Collins (collins@UMBI.UMD.EDU) wrote: > On 10 Jun 1997 afdl@umbi.umd.edu wrote: ^^^^^^^^^^^^^ That's odd. Apparently either your news server or your news reader got confused by the ; in the From: line. See below. > > Check out this site, it has tons of Nude Teenagers... > > ==================================================================== > Attention all subscribers! This offensive message did not come from > anyone at umbi. The "from" address was faked. Is there no way to stop > this sort of thing? -John Collins The best is to complain to postmaster@newsfeed.gte.net. Maybe they have an abuse department. If you complain, include the headers of the message, for example: Path: winx03!uni-erlangen.de!news-nue1.dfn.de!news-mue1.dfn.de! news-stu1.dfn.de!news-kar1.dfn.de!news-was.dfn.de!news.maxwell.syr.edu! europa.clark.net!newsfeed.gte.net!news Message-ID: <5njdak$5vv$3@news9.gte.net> From: afdl;js@;klajsdfals.com Newsgroups: bionet.molbio.proteins Subject: -Hot Teens that want to be fucked Date: 10 Jun 1997 11:20:52 GMT Lines: 12 Organization: GTE Intelligent Network Services, GTE INS NNTP-Posting-Host: 1cust148.tnt1b.lax3.da.uu.net X-Auth: 471D9E0B8F8E5702D6560413 You might also consider complaining to one of the addresses given below. ~% whois gte.net [rs.internic.net] GTE Intelligent Network Services (GTE2-DOM) 5525 MacArthur Blvd. IRVING, TX 75038 USA Domain Name: GTE.NET Administrative Contact: Zinnel, Robert (RZ62) rzinnel@GTE.NET 214-751-3850 Technical Contact, Zone Contact: Ward, Matt (MW837) mward@GTE.NET (972) 751-3851 --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@net.bio.net Wed Jun 11 23:00:00 1997 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.erols.net!newsfeed.nacamar.de!fu-berlin.de!uniol!news.uni-stuttgart.de!news.urz.uni-heidelberg.de!news.dkfz-heidelberg.de!news From: E.Ratsch@DKFZ-Heidelberg.de (Esther Ratsch) Newsgroups: bionet.molbio.proteins Subject: Protein elution from PVDF membrane Date: 12 Jun 1997 12:22:21 GMT Organization: DKFZ Lines: 7 Message-ID: <5noplt$hnr@news.inet.dkfz-heidelberg.de> Reply-To: E.Ratsch@DKFZ-Heidelberg.de NNTP-Posting-Host: atv-alt.inet.dkfz-heidelberg.de X-Newsreader: WinVN 0.92.6+ Hello! Can anybody please provide me with a method to elute protein from a PVDF membrane? Thanks in advance for any help, Esther From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!sas.upenn.edu!fdaldal From: fdaldal@sas.upenn.edu ("Fevzi Daldal") Newsgroups: bionet.molbio.proteins Subject: postdoc Date: 13 Jun 1997 16:32:58 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 29 Sender: daemon@net.bio.net Distribution: world Message-ID: <199706132333.TAA01858@orion.sas.upenn.edu> NNTP-Posting-Host: net.bio.net There is an open position for a Postdoctoral Fellow in our group at the University of Pennsylvania, Department of Biology, Plant Science Institute. The position is effective immediately. The current work is on the structure, function, regulation and biogenesis of cytochrome complexes of photosynthetic bacteria, with emphasis on the molecular genetic and biochemical approaches, as model systems for higher organisms [for our most recent work please see J. Bacteriol. 177: 608-6139 (on cyt cy); Biochemistry 34: 15979-16012 and BBA 1275: 61-69 (on cyt bc1 complex); Biochemistry 33: 3120-3127 (on cyt cbb3 oxidase) and J. Bacteriol 178: 5279-5290 (on cyt c biogenesis)]. The ideal candidate for this position should have a solid background either in bscterial molecular genetics or protein biochemistry and spectroscopy, and a desire to learn multidisciplinary approaches. The salary will be commensurate with experience, and the person interested should send me a CV, description of research accomplishments and references. For additional information I can be reached at (215) 898-4394, Fax: (215) 898-8780 and my Email address is fdaldal@sas.upenn.edu Fevzi Daldal Professor fdaldal@sas.upenn.edu U. Penn/Biology phone:(215) 898-4394 204 Mudd Bldg. fax: (215) 898-8780 Phila PA 19104-6018 University of Pennsylvania Biology Web Site http://www.sas.upenn.edu/biology/ From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!agate!howland.erols.net!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!news.apfel.de!univ-lyon1.fr!cnusc.fr!ciril.fr!grenet.fr!news.imag.fr!macjesior.imag.fr!user From: jean-claude.jesior@imag.fr (J.C. Jesior) Newsgroups: bionet.molbio.proteins Subject: Protein modelling freeware Date: Fri, 13 Jun 1997 14:43:55 +0100 Organization: Institut Albert Bonniot, CNRS Lines: 102 Message-ID: NNTP-Posting-Host: macjesior.imag.fr Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: 8bit ANNOUNCING THE NEW PPC VERSION OF 'FOLDIT (LIGHT) v.4.2.6Ή a molecular modelling freeware for MacOS compatible computers. €€€€€€€€€€€€€€€€€€€€€ FREEWARE €€€€€€€€€€€€€€€€€€€€€ Enhanced PPC version DOWNLOAD * ŒFoldIt (light) v.4.2.6Ή can be downloaded from the following address: ftp://ftp.imag.fr/pub/TIMC/FoldIt.html * ŒFoldIt (light) FilesΉ can also be downloaded from the same address (it contains a set of 20 protein structure files). WHAT 'FOLDIT (LIGHT)' DOES? 'FoldIt (light)' is a molecular modelling program to visualize and manipulate proteins. 'FoldIt (light)' is an interactive program with a user friendly interface. The goal of this program was to build an integrated environment in which statistical analysis as well 3D observations could be realized on PDB files without having to transfer files or swap machines. Our major underlying research project is still to try to improve the folding prediction methods (hence the name 'FoldIt'). This is also the sole desactivated feature in this released version (hence the adjective '(light)'). 'FoldIt (light)' is intented to provide the possibility to analyze proteins up to 1600 residues in size, to visualize and manipulate them interactively. It can directly read any protein coordinate text file from the Brookhaven Protein Data Bank (PDB) (Bernstein et al., 1977) including hetero-atoms and water molecules. 'FoldIt (light)' has two main windows: a color image window to display the protein structure and a text window to record the result of all operations requested by the user. The protein structure can be manipulated easily in real time with the mouse, zoomed or observed in stereo. Structure movement can also occur stepwise for a more precise control. Animations can be created. Steric conflicts, disulfide bonds, hydrogen and ionic interactions can be located and displayed in the protein structure. These interactions are also reported in the text window. Atoms and residues can be tagged individually (or globally) and structural information can then be extracted. Portions of a structure can be read into memory or displayed. Two structures can be read at the same time in memory and can be overlapped automatically. The second structure can be manipulated independently of the first. Bonds can be rotated and atomic parameters can be changed. The sequential folding of a protein can be simulated. It is possible to create a protein de novo from the menu or by entering the sequence from the keyboard. Structures can then be manipulated locally or forced into helices. Site directed mutations can be simulated. The application can process structures in the batch mode to extract a number of structural features: Ramachandran plots, SS-bond plots, H-bond plots. Statistics on atomic parameters are displayed as histograms. The content of image and text windows as well as histograms can be saved to disk. SOFTWARE ENVIRONMENT 'FoldIt (light)' can only work on Macintosh computers running under system 7.0 and higher. It has been developed with Symantec Think Pascal and includes 55000 lines of code. The application can run in the background which is convenient for lengthy statistical procedures. 'FoldIt (light)' does not support printing but can save the content of its text window (report of different operations and data analysis) and the content of the image window (result of graphic operations) in TEXT or PICT formats. These files can then be manipulated by other specialized applications and printed. HARDWARE ENVIRONMENT 'FoldIt (light)' runs only on MacOs compatible computers equipped with a PPC processor. The application itself requires 2 MB of memory; 3.3 KB of additional memory are required for each read residue: this means that 660 KB of extra RAM are necessary to read a typical 200-residue protein. Color monitors are preferred to give a better depth perception but black & white monitors may also be used. DOCUMENTATION On-line help as well as balloon help are supported, and a printable stand alone document is provided. Several small size protein structures are included as example. ****************************************************** FoldIt (light) freeware by Jean-Claude JESIOR / CNRS, Grenoble, France e-mail: jean-claude.jesior@imag.fr Version 4.2.6 - 11 june 1997 developed in Pascal with Symantec Think & Metrowerks Code Warrior. ****************************************************** -- Jean-Claude Jesior jean-claude.jesior@imag.fr From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!agate!spool.mu.edu!uwm.edu!vixen.cso.uiuc.edu!howland.erols.net!newsfeed.internetmci.com!news2.fibr.net!news1.fibr.net!news.fibr.net!news From: Newsgroups: bionet.molbio.proteins Subject: http://www.planetfun.com/videosex.htm Date: 14 Jun 1997 05:49:41 GMT Lines: 3 Message-ID: <5ntbdl$eb3@nimitz.fibr.net> NNTP-Posting-Host: port1-6.lconn.com http://www.planetfun.com/videosex.htm Adults Only From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!daresbury!lyra.csx.cam.ac.uk!news.ox.ac.uk!worf.molbiol.ox.ac.uk!rgrant From: rgrant@see.sig.for.address (Richard P Grant) Newsgroups: bionet.software,bionet.molbio.proteins Subject: Structure similarity search Followup-To: bionet.software Date: 13 Jun 1997 16:06:12 GMT Organization: Oxford University Lines: 20 Message-ID: <5nrr5k$h9c@news.ox.ac.uk> NNTP-Posting-Host: worf.molbiol.ox.ac.uk X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.software:18821 bionet.molbio.proteins:10959 Hi people, Given a protein domain structure (100 amino acids) we would like to search for similar domains in (a) 1 protein (b) the entire sequence databases. The problem is of course that there is no solved structure for (a) and most of (b).... so some form of prediction, threading or modelling will have to be done as part of the search/comparison. Ideas? I can't find exactly what I want on the Web, although the most excellent PredictProtein service in Heidelberg may do what we want. We're looking for either an internet-based resource, or a standalone that will run under Unix or MacOS. Thanks, please followup to bionet.software so that everyone can see if there is a solution. -- Richard P. Grant MA DPhil University of Oxford | rgrant@molbiol http://www.molbiol.ox.ac.uk/~rgrant FFPGP | .ox.ac.uk -----------------------# Trust me - I'm a doctor. #----------------------- From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!CCWF.CC.UTEXAS.EDU!Ldarj From: Ldarj@CCWF.CC.UTEXAS.EDU ("Lavan Dajarnia, Ph.D.") Newsgroups: bionet.molbio.proteins Subject: Please Help: Protein bands' lateral spreading Date: 13 Jun 1997 18:11:00 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <33A1FAA5.347F@ccwf.cc.utexas.edu> Reply-To: ldarj@ccwf.cc.utexas.edu NNTP-Posting-Host: net.bio.net Dear Folks Could anybody help me in solving the problem of protein bands' lateral spreading, which unfortunately occures with my SDS-PAGE ? Can you give main reasons of band lateral diffusion? I work with algae crude protein extracts. Those cells initially were grown at high salt (possible interferring agen), but the cells were washed with a buffer with a very low salt. Anyway gel lane wides at front causing problems in correct gel slicing. Any suggestions are highly appreciated. Thanks and awaiting your responces. Levan darjania From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 13 Jun 1997 02:00:09 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 233 Sender: daemon@net.bio.net Distribution: world Message-ID: <199706130900.CAA03711@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. From owner-proteins@net.bio.net Thu Jun 12 23:00:00 1997 Path: biosci!sas.upenn.edu!fdaldal From: fdaldal@sas.upenn.edu ("Fevzi Daldal") Newsgroups: bionet.molbio.proteins Subject: postdoctoral position Date: 13 Jun 1997 20:10:37 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: <199706140311.XAA15554@orion.sas.upenn.edu> NNTP-Posting-Host: net.bio.net There is an open position for a Postdoctoral Fellow in our group at the University of Pennsylvania, Department of Biology, Plant Science Institute. The position is effective immediately. The current work is on the structure, function, regulation and biogenesis of cytochrome complexes of photosynthetic bacteria, with emphasis on the molecular genetic and biochemical approaches, as mode