From owner-proteins@net.bio.net Fri Dec 01 22:00:00 1995 Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins,bionet.cellbiol,bionet.general Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!nih-csl!postman From: bernard@elsie.nci.nih.gov (Bernard Murray) Subject: Re: HELP! Need alternative to BSA as carrier Message-ID: <1995Dec2.230814.22111@alw.nih.gov> Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: ermintrude.nci.nih.gov Organization: National Institutes of Health, Bethesda, MD X-Newsreader: WinVN 0.99.7 References: Date: Sat, 2 Dec 1995 23:08:14 GMT Lines: 44 Xref: biosci bionet.molbio.methds-reagnts:37110 bionet.molbio.proteins:6395 bionet.cellbiol:3566 bionet.general:18702 In article , fergusb@microbio.lifesci.ucla.edu says... > >WELL KLH IS THE USUSAL OTHER OPTION BUT I DO REMEMBER HEARING ABOUT >SOMEONE WHO USED THE LARGE CARBOHYDRATE DEXTRAN AS A ACARRIER WITH SUCCESS >- SORR, CAN'T REMEMBER THE REFERENCE BUT IT WAS TO DO WITH THE HERBICIDE >ATRAZINE A STHE HAPTEN SO IF YOU SEARCH WITH DEXTRAN, ATRAZINE CARRIER >HAPTEN YOU SHOULD FIND IT - ACTUALLY, IT MAY WELL HAVE BEEN IN THE JOURNAL >OF IMMUNOLOGICALO METHODS > >GOOD LUCK > >FERGUS >UCLA There's no need to shout! >In article , >michael.didonato@utoronto.ca (Michael DiDonato) wrote: > >> I am desperately looking for an alternative to using BSA or any other >> albumins as a carrier protein in cell culture experiments. Any >> help/advice would be extremely appreciated. >> >> Mike >> Michael DIDonato >> michael.didonato@utoronto.ca Can you give a little more information? I interpreted your question as a need for carriers for solubilising substances to be added to tissue culture cells (eg. many peptide hormones are dissolved in solutions containing BSA before addition to cultured cells). In this case I would suggest gelatin as an alternative but I've heard of dextran or polyvinylpyrolidine being used. If, indeed, you are asking for an immunological carrier for conjugation then I second KLH and would maybe also suggest lysozyme or a globulin (eg. lactoglobulin) but my experience is only with in vivo immunisation. What property of BSA is it that you don't like? Good luck, Bernard Bernard Murray, Ph.D. bernard@elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA) From owner-proteins@net.bio.net Fri Dec 01 22:00:00 1995 Path: biosci!newshost.lanl.gov!ncar!news.internetMCI.com!newsfeed.internetmci.com!in2.uu.net!interaccess!d129.w.interaccess.com!wpenrose From: wpenrose@interaccess.com (William R. Penrose) Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: Sat, 2 Dec 1995 20:58:20 Organization: InterAccess,Chicagoland's Full Service Internet Provider Lines: 29 Message-ID: References: NNTP-Posting-Host: d129.w.interaccess.com X-Newsreader: Trumpet for Windows [Version 1.0 Rev B final beta #4] Xref: biosci bionet.general:18703 bionet.molbio.proteins:6396 bionet.population-bio:1685 sci.chem:44219 In article Bob_Hoesch@fws.gov (Bob Hoesch) writes: >If one is using hemoglobin as a marker for analyzing population >genetics, a standard procedure is to treat the blood sample with low >concentrations of cyanide prior to IEF (isoelectric focusing). This >results in the elimination of "spurious" bands, and apparently makes >the resulting banding patterns amenable to genetic analysis. I've heard >this procedure described as "reduction of methemoglobin" (reduction of >the oxidized iron back to the reduced state), but this doesn't make sense >to me in terms of the chemistry. Can someone explain what is happening in >this cyanide-induced reduction in the number of hemoglobin bands? How could >cyanide reduce Fe3+ to Fe2+? It doesn't. Methemoglobin forms a tight complex with CN, and ordinary Hb doesn't. Presumably this affects the absorption bands enough to eliminate them as interference. One of the treatments for HCN poisoning is to inhale an organic nitrite, which oxidizes Hb to mHb. The CN is then tied up by the mHb instead of trashing the cytochroma oxidase, which is its toxic target. ******************************************************** Bill Penrose, Transducer Research, 600 North Commons Dr., Suite 117, Aurora, IL 60504, 708-978-8802, fax -8854. Email wpenrose@interaccess.com The Contract on America: Free markets for the middle class, socialism and subsidies for the rich. Vote Republican. ******************************************************** From owner-proteins@net.bio.net Fri Dec 01 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!info.ucla.edu!library.ucla.edu!news.ucdavis.edu!dcn71.dcn.davis.ca.us!user From: ijiwaru@wheel.dcn.davis.ca.us Newsgroups: bionet.molbio.proteins Subject: Re: protein extraction Date: Fri, 01 Dec 1995 23:28:13 +0500 Organization: University of California, Davis Lines: 27 Distribution: world Message-ID: References: <19951201114156858.savidge@unb.ca> NNTP-Posting-Host: dcn71.dcn.davis.ca.us X-Newsreader: Value-Added NewsWatcher 2.0b27.1+ How about biotinylating your monoclonal and dispensing with the secondary Ab? Just out of curiousity, how are you going to get the mAb across the membrane? liposome fusion? L In article <19951201114156858.savidge@unb.ca>, savidge@unb.ca wrote: > We're thinking about using a monoclonal antibody (IgG) in an > attempt to immunolocalize a water-soluble molecule (MW <200) that > presumably exists in the cytosol or vacuole of plant cells. We think we > can get the McAb into the cells OK, then cross-link the McAb-hapten > complex using a fixative to another protein (or other structure), and > follow with a secondary anti-IgG `reporter' antibody. We know that > we cannot fix the hapten prior to introducing the McAb because it would > make the epitope unrecognizable. Washing away unbound secondary antibody > should not be a problem; however, I can't think how - prior to the > cross-linking step - we can wash away unbound primary McAb without > concomitantly losing the McAb-hapten complex. If anybody has experience > with this I'd appreciate receiving advice. > ********************************************************************** > Rod Savidge, PhD, Professor | E-mail: savidge@unb.ca > Faculty of Forestry and \|/ > Environmental Management \ | / Phone: (506) 453-4919 > University of New Brunswick _\/ | \/_ > Fredericton, NB CANADA \|/ Fax: (506) 453-3538 > E3B 6C2 | From owner-proteins@net.bio.net Sat Dec 02 22:00:00 1995 Path: biosci!galaxy.ucr.edu!usenet From: Protic Channel Newsgroups: bionet.molbio.proteins Subject: (none) Date: Sun, 03 Dec 1995 04:02:59 +0000 Organization: University of California, Riverside Lines: 4 Message-ID: <30C12173.2C52@protic.channel.sci> NNTP-Posting-Host: a-ilab2.ucr.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b1 (Macintosh; I; PPC) Please give your opinions on protic channels. Reply on anything about them, whether you think they exist or not, or whether you think they can be cloned or not, etc. Thank you. From owner-proteins@net.bio.net Sat Dec 02 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!cs.utexas.edu!geraldo.cc.utexas.edu!bashful.cc.utexas.edu!hantash From: hantash Newsgroups: bionet.molbio.proteins Subject: Hydropathy programs !! Date: Sun, 3 Dec 1995 10:56:29 -0600 Organization: The University of Texas at Austin, Austin, Texas Lines: 9 Message-ID: NNTP-Posting-Host: bashful.cc.utexas.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Hi; Does anyone know of a paper and /or a program to predict proteins that may have hydropathic domains ? Thanks Feras hantash@ccwf.cc.utexas.edu From owner-proteins@net.bio.net Sat Dec 02 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!cs.utexas.edu!geraldo.cc.utexas.edu!bashful.cc.utexas.edu!hantash From: hantash Newsgroups: bionet.molbio.proteins Subject: Also Amphipathic Helices ? Date: Sun, 3 Dec 1995 13:06:37 -0600 Organization: The University of Texas at Austin, Austin, Texas Lines: 9 Message-ID: NNTP-Posting-Host: bashful.cc.utexas.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Hi I forgot to ask also about papers and /or programs that could predict amphipathic helices in a protein. Any help would be much appreciated. Thanks Feras hantash@ccwf.cc.utexas.edu From owner-proteins@net.bio.net Sat Dec 02 22:00:00 1995 Path: biosci!agate!cocoa.brown.edu!cis-ts6-slip11.cis.brown.edu!user From: Stephen_Lasky@brown.edu (Stephen R. Lasky) Newsgroups: bionet.molbio.proteins Subject: Re: Hydropathy programs !! Date: Sun, 03 Dec 1995 13:21:33 -0400 Organization: Roger Williams Medical Center/Brown University Lines: 29 Message-ID: References: NNTP-Posting-Host: cis-ts7-slip2.cis.brown.edu X-Newsreader: Value-Added NewsWatcher 2.0b24.0+ In article , hantash wrote: > Hi; > > Does anyone know of a paper and /or a program to predict proteins that > may have hydropathic domains ? > > Thanks > > Feras > hantash@ccwf.cc.utexas.edu Two papers describe the commonly used algorithms: Hopp & Woods, and Kyte & Doolittle. They are back around 1980 or 1981. I think MacVector lets you choose which hydropathicity algorithm you want to use. I'm sure that most of the protein analysis software does similar analyses (GeneWorks, GCG, etc). SRLasky -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Stephen R. Lasky Ph.D. Brown U/Roger Williams Medical Center, Providence, RI. Phone: 401-456-5672 Fax: 401-456-6569 e:mail: Stephen_Lasky@brown.edu =================================================================== America may be unique in being a country which has leapt from barbarism to decadence without touching civilization. John O'Hara. ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!torn!utnut!oci!news From: Flip Hoedemaeker Subject: Re: Also Amphipathic Helices ? Content-Type: multipart/mixed; boundary="---------------------------145201218724890" Message-ID: <30C36C1F.21A3@oci.utoronto.ca> Sender: news@oci.utoronto.ca Organization: The Holistic Detective Agency References: Mime-Version: 1.0 Date: Mon, 4 Dec 1995 21:46:07 GMT X-Mailer: Mozilla 2.0b2a (Windows; I; 16bit) Lines: 30 This is a multi-part message in MIME format. -----------------------------145201218724890 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit This is available in the GCG package: HELICALWHEEL HelicalWheel plots a peptide sequence as a helical wheel to help you recognize amphiphilic regions. Its probably just as easy to do it by hand, as soon as you know where your helices might be... Flip -----------------------------145201218724890 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Content-Disposition: inline; filename="1.TXT" HELICALWHEEL HelicalWheel plots a peptide sequence as a helical wheel to help you recognize amphiphilic regions. -----------------------------145201218724890-- From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!tank.news.pipex.net!pipex!soap.news.pipex.net!pipex!usenet From: M J Geisow Newsgroups: bionet.molbio.proteins Subject: STRUCTURAL BIOLOGY WEB UPDATES Date: 5 Dec 1995 02:07:29 GMT Organization: BIODIGM Ltd. Lines: 26 Message-ID: <4a09h1$8g9@soap.news.pipex.net> NNTP-Posting-Host: ai137.du.pipex.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1 (Windows; I; 16bit) STRUCTURAL BIOLOGY UPDATES 1. Calendar of events updated: http://www.cryst.bbk.ac.uk/CEC/calendar.html 2. Posters at 5th International Perspectives on Protein Engineering conference now each get a mini - home page! Poster prizes now offered (UK Biochemical Society) http://www.cryst.bbk.ac.uk/CEC/5posters.html 3. Informal biotech / pharma company contact service open at the above conference. Please contact me if your company is interested (in return for meeting sponsorship) http://www.cryst.bbk.ac.uk/CEC/contact.html 4. (UK citizen) The BBSRC has announced a second call for bioinformatics outline proposals (closing 9 Feb 96) http://www.cryst.bbk.ac.uk/CEC/key.htm -- BIODIGM Tel: +44 (0) 1949 839077 Fax: +44 (0) 1949 831886 Email: biodigm@dial.pipex.com From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bloom-beacon.mit.edu!newsfeed.internetmci.com!vixen.cso.uiuc.edu!uchinews!news From: Kelly Ambler Subject: Re: Disappearing SDS-PAGE bands X-Nntp-Posting-Host: hearts-pc45.bsd.uchicago.edu Content-Type: text/plain; charset=us-ascii Message-ID: Sender: news@midway.uchicago.edu (News Administrator) Content-Transfer-Encoding: 7bit Organization: University of Chicago -- Academic Computing Services References: <30C36A66.564B@oci.utoronto.ca> Mime-Version: 1.0 Date: Tue, 5 Dec 1995 00:58:58 GMT X-Mailer: Mozilla 1.1N (Windows; I; 16bit) Lines: 18 I don't think it was the staining conditions - I ran, and thus stained, different individual gels (all polymerized in one batch) on separate days, yet had the same band pattern in all the gels. I reuse my staining solution and did not have this problem with other gels stained at later date. These were 11% acrylamide gels. One other fact - the demarcation between bands and no-bands was quite sharp at the bottom of the "problem zone", but appeared to fade out gradually at the top of the "problem zone". I have talked to several people about this problem, and we're all quite puzzled. Mostly, I would like to ensure that it doesn't occur again! Kelly Ambler University of Chicago From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!pravda.aa.msen.com!nntp.coast.net!torn!resunix.ri.sickkids.on.ca!NewsWatcher!user From: michael.didonato@utoronto.ca (Michael DiDonato) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins,bionet.cellbiol,bionet.general Subject: Re: HELP! Need alternative to BSA as carrier Date: 4 Dec 1995 17:07:50 GMT Organization: The Hospital for Sick Children Lines: 8 Message-ID: References: <49nvht$741c@tigger.cc.uic.edu> NNTP-Posting-Host: 142.20.5.141 Xref: biosci bionet.molbio.methds-reagnts:37150 bionet.molbio.proteins:6404 bionet.cellbiol:3585 bionet.general:18718 In article <49nvht$741c@tigger.cc.uic.edu>, Keld Sorensen wrote: > If you mean "carrier protein" for antibody production, Actually I need it for cell culture not antibody production. Thanks. Mike From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!pravda.aa.msen.com!nntp.coast.net!torn!resunix.ri.sickkids.on.ca!NewsWatcher!user From: michael.didonato@utoronto.ca (Michael DiDonato) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins,bionet.cellbiol,bionet.general Subject: Re: HELP! Need alternative to BSA as carrier Date: 4 Dec 1995 17:06:59 GMT Organization: The Hospital for Sick Children Lines: 12 Message-ID: References: NNTP-Posting-Host: 142.20.5.141 Xref: biosci bionet.molbio.methds-reagnts:37149 bionet.molbio.proteins:6403 bionet.cellbiol:3584 bionet.general:18717 ACTUALLY, IT MAY WELL HAVE BEEN IN THE JOURNAL > OF IMMUNOLOGICALO METHODS > > GOOD LUCK > > FERGUS > UCLA I actually meant that I needed a carrier to use in tissue culture experiments, not for antibody production. Thanks anyway. Mike From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.kei.com!ub!newserve!rebecca!pauling.wadsworth.org!tivol From: tivol@news.wadsworth.org (William Tivol) Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Followup-To: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Date: 1 Dec 1995 22:29:47 GMT Organization: Wadsworth Center, NY Health Dept. Lines: 15 Message-ID: <49nvkr$q0u@pauling.wadsworth.org> References: NNTP-Posting-Host: alcor.wadsworth.org X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.general:18716 bionet.molbio.proteins:6402 bionet.population-bio:1687 sci.chem:44293 Bob Hoesch (Bob_Hoesch@fws.gov) wrote: : If one is using hemoglobin as a marker for analyzing population : genetics, a standard procedure is to treat the blood sample with low : concentrations of cyanide prior to IEF (isoelectric focusing). This : results in the elimination of "spurious" bands, and apparently makes : the resulting banding patterns amenable to genetic analysis. I've heard : this procedure described as "reduction of methemoglobin" (reduction of : the oxidized iron back to the reduced state), but this doesn't make sense : to me in terms of the chemistry. Can someone explain what is happening in : this cyanide-induced reduction in the number of hemoglobin bands? How could : cyanide reduce Fe3+ to Fe2+? Dear Bob, Just a thought, but Fe3+ + CN- = Fe(2+)CN. Yours, Bill Tivol From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.internetMCI.com!darwin.sura.net!maze.dpo.uab.edu!usenet From: "N.Ambalavanan MD" Newsgroups: bionet.molbio.proteins Subject: Cross reactivity between Porcine EGF and Human EGF on ELISA? Date: Sun, 03 Dec 1995 19:11:50 -0600 Organization: UAB Lines: 6 Message-ID: <30C24AD6.2DD5@uabdpo.dpo.uab.edu> NNTP-Posting-Host: tty13.maze.ppp.uab.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b2 (Windows; I; 32bit) Hi! I will be trying to determine the magnitude of production of EGF production by porcine cells using an ELISA kit designed for human EGF ( R&D systems) , since there is no kit for the porcine form, and my attempts at designing an ELISA myself were a flop. Does anyone have any idea how much porcine EGF is similar to human EGF, and if they crossreact on ELISA using monoclonals? From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!torn!utnut!oci!news From: Flip Hoedemaeker Subject: Re: Disappearing SDS-PAGE bands Content-Type: text/plain; charset=us-ascii Message-ID: <30C36A66.564B@oci.utoronto.ca> Sender: news@oci.utoronto.ca Content-Transfer-Encoding: 7bit Organization: The Holistic Detective Agency References: Mime-Version: 1.0 Date: Mon, 4 Dec 1995 21:38:46 GMT X-Mailer: Mozilla 2.0b2a (Windows; I; 16bit) Lines: 17 Kelly Ambler wrote: > I recently had some very strange results from my Coumassie stained > gels. The proteins in the middle of the gels "disappeared". The > bands at the bottom half of the gel and at the very top of the gel > appeared to run as usual. Very strange indeed! When I do electrophoresis of cell lysates I some- times see a lack of bands around the 15-20 kD range, but nothing as serious as your gels. Have you tried restaining your gels? Maybe your staining procedure is not optimal, i.e. the solution is too dilute or the time too short. The middle of the gel we be the last to stain properly... Do you use gradient gels? Flip From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!usc!news.cerf.net!news From: "David R. Higgins" Newsgroups: bionet.molbio.proteins Subject: Pichia meeting Date: Mon, 04 Dec 1995 10:02:58 -0800 Organization: CERFnet Lines: 8 Message-ID: <30C337D2.4BA8@powergrid.electriciti.com> Reply-To: dhiggins@powergrid.electriciti.com NNTP-Posting-Host: 199.106.180.163 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b1 (Macintosh; I; PPC) Current Topics in Gene Expression: Pichia pastoris". Meeting on heterologous protein expression in Pichia pastoris will be held in San Diego March 3-6, 1996. Sessions include Pichia Cell Biology, Post-translation modifications/protein processing, Expression optimization, scale-up/fermentation, Industrial applications, and New developments in expression technology. The meeting is being hosted by Invitrogen and RCT. Registration deadline is February 9th. For more information please send reply e-mail to From owner-proteins@net.bio.net Sun Dec 03 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: kakoepli@aol.com (KAKOEPLI) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins,bionet.cellbiol,bionet.general Subject: Re: HELP! Need alternative to BSA as carri Date: 4 Dec 1995 22:44:54 -0500 Organization: America Online, Inc. (1-800-827-6364) Lines: 16 Sender: root@newsbf02.news.aol.com Message-ID: <4a0f7m$59n@newsbf02.news.aol.com> References: Reply-To: kakoepli@aol.com (KAKOEPLI) NNTP-Posting-Host: newsbf02.mail.aol.com Xref: biosci bionet.molbio.methds-reagnts:37170 bionet.molbio.proteins:6410 bionet.cellbiol:3591 bionet.general:18727 In certain cell culture systems liposomes are substituted for albumin or serum. These serum free media can be used to evaluate the effect of protein (albumin or other) binding on cell-uptake of drugs and other molecules. Descriptions of serum free media are available in the literature. Often cells need to be weaned off the serum in steps for adequate viability in the serum free media. Viability probably depends on cell type and other variables. Note that chemicals/nutrients present in the medium (eg, fatty acids) can be dramatically more cytotoxic to the cells due to increased cell uptake in the absence of an extracellular binding protein which normally "buffers" the free level available to the cells. KAKOEPLI@aol.com From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!fdn.fr!jussieu.fr!jouy.inra.fr!usenet From: villatte@versailles.inra.fr (François Villatte) Newsgroups: bionet.molbio.proteins Subject: glycosylation Date: 5 Dec 1995 10:12:06 GMT Organization: INRA Phyto Lines: 5 Message-ID: <4a15tm$krn@saphir.jouy.inra.fr> NNTP-Posting-Host: 147.99.4.20 X-Newsreader: WinVN 0.91.6 Hello, I currently search a colorimetric method to detect glycoproteins on a native PAGE. Moreover, I would like to know if a GST detection method on PAGE exist. Francois From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!ihnp4.ucsd.edu!news.service.uci.edu!design.eng.uci.edu!pecota From: Doug Pecota Newsgroups: bionet.molbio.proteins Subject: Plasmid R1 Date: Tue, 5 Dec 1995 17:24:11 -0800 Organization: University of California, Irvine Lines: 12 Message-ID: NNTP-Posting-Host: design.eng.uci.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII I am looking for plasmid R1. I called ATCC they said that they do not have any stains containing R1. I contacted several companies that sell plasmids but they did not sell R1. R1 is the mother of many plasmids used in genetic engineering as result many modified R1 type plasmids are available. However for my research I would prefer the R1 wild type plasmid or something very close. Please contact me at pecota@eng.uci.edu if you can give me the plasmid or a source from which I can obtain it. Thanks Doug P. University of Califoria Irvine From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!agate!lhom From: lhom@nature.Berkeley.EDU (Louis Hom) Newsgroups: bionet.molbio.proteins Subject: Protein Models & CAD/CAM Date: 6 Dec 1995 01:32:30 GMT Organization: Agricultural and Resource Economics Lines: 10 Message-ID: <4a2rre$alp@agate.berkeley.edu> NNTP-Posting-Host: nature.berkeley.edu It seems to me that life for some folks would be easier if you could have your computer create a real 3D representation of a protein structure. I know that computer-controlled generation of prototypes in manufacturing is done all the time. Does anyone know of someone trying to apply computer aided manufacturing technology to protein structures? -- _______________________________________________________________________________ Lou Hom >K '93 "Pe--li--gro . . . Pe-li-gro. . . lhom@nature.berkeley.edu Peligro. Ah, peligro!" http://www.ocf.berkeley.edu/~lhom **SPLAT!** From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!tank.news.pipex.net!pipex!acorn!acorn!uknet!ftel.co.uk!bham!bhamcs!news.ox.ac.uk!springer From: springer@immsvr.jr2.ox.ac.uk. (springer) Newsgroups: bionet.molbio.proteins Subject: IEF and conformational change Date: 5 Dec 1995 12:30:00 GMT Organization: Oxford University Lines: 14 Message-ID: <4a1e08$ie3@news.ox.ac.uk> NNTP-Posting-Host: immmac5.jr2.ox.ac.uk X-Posted-From: InterNews 1.0@news.ox.ac.uk. It should be possible to look at the conformational change of a protein (receptor or enzyme) using isoelectric focusing, as the pKs of the amino acid side chain functional groups depend on their chemical environment. Is anybody aware of a study in which this has been done, or some theoretical background? We have a peptide receptor that upon the binding of a (neutral) peptide shifts its isoelectric point, and we are wondering whether this could be attributed to conformational change. Advice would be highly appreciated! Sebastian Springer springer@vax.ox.ac.uk From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!interaccess!d209.nb.interaccess.com!wpenrose From: wpenrose@interaccess.com (William R. Penrose) Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: Tue, 5 Dec 1995 16:51:43 Organization: InterAccess,Chicagoland's Full Service Internet Provider Lines: 39 Message-ID: References: NNTP-Posting-Host: d209.nb.interaccess.com X-Newsreader: Trumpet for Windows [Version 1.0 Rev B final beta #4] Xref: biosci bionet.general:18756 bionet.molbio.proteins:6419 bionet.population-bio:1690 sci.chem:44399 In article dyanega@denr1.igis.uiuc.edu (Doug Yanega) writes: >> them as interference. One of the treatments for HCN poisoning is to >inhale an >> organic nitrite, which oxidizes Hb to mHb. The CN is then tied up by the mHb >> instead of trashing the cytochroma oxidase, which is its toxic target. >This raises a question: I've been told by a colleague that cyanide builds >up in the body tissues over time, and that after years of slight sublethal >doses, one more tiny dose can "push a person over the edge" suddenly. That's a new one on me. But then toxicology has a lot of folklore in it. In fact, the idea of using amyl nitrite as an antidote is a bit of folklore, since if you get a good faceful of HCN you will be off the planet before anyone can find the stuff. The main purpose of amyl nitrite ampoules is probably to make people feel more comfortable working with cyanide. During my undergraduate days, I got a little sniff of HCN and it was like being hit in the face with a two-by-four. It reduced me to semiconsciousness in a second or two, but I recovered over the next few minutes and did not even need treatment. In favor of your friend, there was the case of the silver-recovery plant in Chicago a while ago where a man died after years of leaning over vats containing cyanide used for leaching silver from photo film. The plant owners had criminal charges brought against them. Either the guy died from the cumulative dose, or one day he just got too big a whiff. Perhaps the body cannot replace or regenerate the cytochrome oxidase fast enough to support the attrition from continual exposure? ************************************************************************ Bill Penrose, Sr. Scientist, Transducer Research, Inc., 600 N. Commons Dr, Ste. 117, Aurora IL 60540, 708-978-8802, fax -8854 email wpenrose@interaccess.com "In any field, it is easy to see who the pioneers are -- they are the ones lying face down with arrows in their backs." (Anon.) ************************************************************************ From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!math.ohio-state.edu!magnus.acs.ohio-state.edu!csn!borcore.usbr.gov!sacsa3.mp.usbr.gov!fwssun1.irm.r6.fws.gov!ash.lab.r1.fws.gov!bob!Bob_Hoesch From: Bob_Hoesch@fws.gov (Bob Hoesch) Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: Tue, 5 Dec 1995 13:30:52 Organization: U.S. Fish & Wildlife Service Forensics Laboratory Lines: 50 Message-ID: References: NNTP-Posting-Host: bob.lab.r1.fws.gov X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Xref: biosci bionet.general:18754 bionet.molbio.proteins:6418 bionet.population-bio:1689 sci.chem:44398 Bob Hoesch Wrote: >>>If one is using hemoglobin as a marker for analyzing population >>>genetics, a standard procedure is to treat the blood sample with low >>>concentrations of cyanide prior to IEF (isoelectric focusing). This >>>results in the elimination of "spurious" bands, and apparently makes >>>the resulting banding patterns amenable to genetic analysis. I've heard >>>this procedure described as "reduction of methemoglobin" (reduction of >>>the oxidized iron back to the reduced state), but this doesn't make sense >>>to me in terms of the chemistry. Can someone explain what is happening in >>>this cyanide-induced reduction in the number of hemoglobin bands? How >>>could cyanide reduce Fe3+ to Fe2+? ???wrote >> It doesn't. Methemoglobin forms a tight complex with CN, and ordinary Hb >> doesn't. Presumably this affects the absorption bands enough to eliminate >> them as interference. One of the treatments for HCN poisoning is to >inhale an >> organic nitrite, which oxidizes Hb to mHb. The CN is then tied up by the mHb >> instead of trashing the cytochroma oxidase, which is its toxic target. Doug Yanega wrote: >This raises a question: I've been told by a colleague that cyanide builds >up in the body tissues over time, and that after years of slight sublethal >doses, one more tiny dose can "push a person over the edge" suddenly. This >was part of his mandatory education working in an analytical chemistry lab >some 40 years ago. This seems completely at odds with my understanding of >the chemistry involved in cyanide toxicity, and I'm curious as to whether >this is at all possible - I'm a professional entomologist, and am exposed >to sublethal doses of cyanide hundreds of times a year..... Hoesch writes again: Two comments. 1) If cyanide preferentially binds to metHb, how would this affect the electrophoretic pattern? In practice, one sees a reduction in the number of Hb bands. Could the binding of one CN- per molecule knock the pI of the resulting complex completely out of the gradient? (This is the only thing I can think of) 2) Doug, I've never heard of "cumulative cyanide toxicity" and it doesn't make sense to me either. Cyanide is very water soluble and would tend to be excreted. And if it does bind irreversibly to metHb, the mean lifetime of red blood cells is only something like 120 days, after which they are taken out of circulation. At that point the cyanide should be released and excreted. Bob Hoesch U.S. Fish & Wildlife Forensics Lab From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.sover.net!lori.albany.net!pm1ip25-albny.albany.net!user From: mdeziel@albany.net (Mark R. Deziel, Ph.D.) Newsgroups: bionet.molbio.proteins Subject: Hela Cell 2-d map? Date: Tue, 05 Dec 1995 15:25:45 -0500 Organization: Albany Medical College Lines: 11 Message-ID: NNTP-Posting-Host: pm1ip25-albny.albany.net Could anyone point me to a good annotated map of hela cell proteins separated by 2-d electrophoresis? A Web site would be ideal, but hard copy would be OK too. Thanks, Mark R. Deziel, Ph.D. Assistant Professor of Medicine and Physiology Albany Medical College From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!lll-winken.llnl.gov!uwm.edu!vixen.cso.uiuc.edu!catalpa.inhs.uiuc.edu!user From: dyanega@denr1.igis.uiuc.edu (Doug Yanega) Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: 5 Dec 1995 17:24:51 GMT Organization: Illinois Natural History Survey Lines: 39 Message-ID: NNTP-Posting-Host: catalpa.inhs.uiuc.edu X-Newsreader: Value-Added NewsWatcher 2.0b27.1+ Xref: biosci bionet.general:18743 bionet.molbio.proteins:6416 bionet.population-bio:1688 sci.chem:44389 > In article Bob_Hoesch@fws.gov (Bob Hoesch) writes: > > >If one is using hemoglobin as a marker for analyzing population > >genetics, a standard procedure is to treat the blood sample with low > >concentrations of cyanide prior to IEF (isoelectric focusing). This > >results in the elimination of "spurious" bands, and apparently makes > >the resulting banding patterns amenable to genetic analysis. I've heard > >this procedure described as "reduction of methemoglobin" (reduction of > >the oxidized iron back to the reduced state), but this doesn't make sense > >to me in terms of the chemistry. Can someone explain what is happening in > >this cyanide-induced reduction in the number of hemoglobin bands? How could > >cyanide reduce Fe3+ to Fe2+? > > It doesn't. Methemoglobin forms a tight complex with CN, and ordinary Hb > doesn't. Presumably this affects the absorption bands enough to eliminate > them as interference. One of the treatments for HCN poisoning is to inhale an > organic nitrite, which oxidizes Hb to mHb. The CN is then tied up by the mHb > instead of trashing the cytochroma oxidase, which is its toxic target. This raises a question: I've been told by a colleague that cyanide builds up in the body tissues over time, and that after years of slight sublethal doses, one more tiny dose can "push a person over the edge" suddenly. This was part of his mandatory education working in an analytical chemistry lab some 40 years ago. This seems completely at odds with my understanding of the chemistry involved in cyanide toxicity, and I'm curious as to whether this is at all possible - I'm a professional entomologist, and am exposed to sublethal doses of cyanide hundreds of times a year, virtually every time I use a killing jar. I can't believe that I'm just cranking up some invisible physiological ratchet, and priming myself to drop dead some day when I catch that one fatal whiff. Thanks, -- Doug Yanega Illinois Natural History Survey, 607 E. Peabody Dr. Champaign, IL 61820 USA (217) 244-6817 affiliate, University of Illinois Dept. of Entomology "There are some enterprises in which a careful disorderliness is the true method" - Herman Melville, Moby Dick From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!MANI.CBS.UMN.EDU!npv From: npv@MANI.CBS.UMN.EDU (Nora Plesofsky-Vig) Newsgroups: bionet.molbio.proteins Subject: re: disappearing SDS-PAGE bands Date: 5 Dec 1995 09:06:22 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <9512051702.AA00814@mani.cbs.umn.edu> Reply-To: nora@biosci.cbs.umn.edu NNTP-Posting-Host: net.bio.net I also was having a problem with a particular region of certain gel lanes registering no protein bands. This seems to have been due to some discontinuity in contact between the glass plates used for the gel. When I substituted the glass plates, the problem disappeared (rather than the bands). I don't know if this is the source of your problem or not. It is possible that some component in your system is not well aligned for optimal contact. Nora Plesofsky-Vig nora@biosci.cbs.umn.edu From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!newsflash.concordia.ca!news.mcgill.ca!NewsWatcher!user From: takayesu@medcor.mcgill.ca (Tak) Newsgroups: bionet.molbio.proteins Subject: Re: Gel-elution Date: Tue, 05 Dec 1995 11:29:26 -0500 Organization: McGill University Lines: 6 Message-ID: References: <1995Nov27.134900@crick.rz-berlin.mpg.de> <49g6vh$15@vixen.cso.uiuc.edu> NNTP-Posting-Host: 132.206.101.175 The reference for disulfide crosslinked polyacrylamide gels is : S. Ghaffari et al. in Analytical Biochemistry 171, 352(1988) Isolation of Proteins for Peptide Mapping etc. Tak From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!EU.net!Portugal.EU.net!news.rccn.net!scsing.switch.ch!swidir.switch.ch!in2p3.fr!univ-lyon1.fr!jussieu.fr!jouy.inra.fr!usenet From: villatte@versailles.inra.fr (François Villatte) Newsgroups: bionet.molbio.proteins Subject: glyco Date: 5 Dec 1995 16:09:05 GMT Organization: INRA Phyto Lines: 6 Message-ID: <4a1qr1$247@saphir.jouy.inra.fr> NNTP-Posting-Host: 147.99.4.20 X-Newsreader: WinVN 0.91.6 Hello, I currently search a colorimetric method to detect glycoproteins on a native PAGE. Moreover, I would like to know if a GST detection method on PAGE exist. Thanks. Francois From owner-proteins@net.bio.net Mon Dec 04 22:00:00 1995 Path: biosci!SPARTAN.AC.BROCKU.CA!jatkin From: jatkin@SPARTAN.AC.BROCKU.CA (Jeffrey Atkinson) Newsgroups: bionet.molbio.proteins Subject: metal vs PEEK flow path in protein HPLC Date: 5 Dec 1995 08:19:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 38 Sender: daemon@net.bio.net Distribution: world Message-ID: <199512051610.LAA16245@spartan.ac.BrockU.CA> NNTP-Posting-Host: net.bio.net Protein Chromatography Experts: I am in the process of purchasing a new HPLC which will be used to detect and purify proteins and peptides as well as more "normal" small organic molecules. I have done this kind of work in the past, quite efficiently, on what would now be called normal systems, i.e. the old stainless steel flow paths. Manufacturers now make a big deal of offering biocompatible flow paths in inert polymers such as PEEK, and titanium systems that are supposed to lower the contamination of protein samples with metal ions. 1) When does metal ion contamination become a problem? Does it interfer with normal sequencing chemistry? (I suspect it shouldn't) Or is it more important for maintaining biological activity? 2) Will PEEK systems still be reliable in the long run with organic solvents? I would assume that polar solvents such as MeOH, MeCN, etc would be OK during RP-HPLC but that noraml phase solvents such as CH2Cl2 and hexane would be a problem. Any other hints or concerns with either choice would be greatly appreciated. Jeffrey Dr. Jeffrey Atkinson Department of Chemistry Brock University St.Catharines, Ontario Canada L2S 3A1 jatkin@spartan.ac.BrockU.ca http://chemiris.labs.brocku.ca/staff/atkinson/atkinson.html phone: 905 688-5550 ext 3967 fax: 905 682-9020 905 688-2789 From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!EU.net!peer-news.britain.eu.net!zippy.dct.ac.uk!dundee.ac.uk!dundee.ac.uk!not-for-mail Newsgroups: bionet.molbio.proteins Subject: Re: glyco Message-ID: <4a4325$cno@dux.dundee.ac.uk> From: hjtreuma@dux.dundee.ac.uk (H.J. Treumann Biochemistry ext 4301) Date: 6 Dec 1995 12:41:41 -0000 References: <4a1qr1$247@saphir.jouy.inra.fr> Organization: The University, Dundee, DD1 4HN, Scotland, UK. NNTP-Posting-Host: dux.dundee.ac.uk X-Newsreader: TIN [version 1.2 PL1] Lines: 12 François Villatte (villatte@versailles.inra.fr) wrote: : Hello, : I currently search a colorimetric method to detect glycoproteins on a : native PAGE. : Moreover, I would like to know if a GST detection method on PAGE exist. : Thanks. : Francois -- Achim Treumann tel. +44-1382-344301 Carbohydrate Research Centre fax +44-1382-322583 Department of Biochemistry Dundee DD1 4HN, Scotland, UK email a.treumann@dundee.ac.uk From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!csn!nntp-xfer-2.csn.net!csn!ub!acsu.buffalo.edu!ubvms.cc.buffalo.edu!v059lr6y From: v059lr6y@ubvms.cc.buffalo.edu (Timothy E Berlinski) Newsgroups: bionet.molbio.proteins Subject: Extraction help Date: 6 Dec 1995 11:18 EST Organization: University at Buffalo Lines: 7 Distribution: world Message-ID: <6DEC199511182884@ubvms.cc.buffalo.edu> NNTP-Posting-Host: ubvmsb.cc.buffalo.edu News-Software: VAX/VMS VNEWS 1.41 I know this has nothing to do with protein but I don't have time to spend 2 hrs looking for a group that does. O.K. I was wondering if anybody has a step wise method for extracting DNA and RNA from mammary cells. It probably doesn't have to be mammary cells in particular. Tim Berlinski From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!UQTR.UQuebec.ca!Mario_Fragata From: Mario_Fragata@UQTR.UQuebec.ca Newsgroups: bionet.molbio.proteins Subject: Re: NATO-ASI on Biomolecular structure and dynamics"NATO Advanced Study Institute" Date: 6 Dec 1995 10:22:06 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <9512061820.AA69224@neptune.UQTR.UQuebec.ca> NNTP-Posting-Host: net.bio.net Dear Prof. Vergoten, I would be very much obliged to you if you could send me detailed information on the symposium "Biomolecular structure ...:" Best regards Mario Fragata Universite du Quebec a Trois-Rivieres Departement de chimie et biologie, Section de chimie Trois-Rivieres, Que, G9A 5H7, Canada tel 819-376-5077 fax 819-376-5057 email fragata@uqtr.uquebec.ca From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!EU.net!peer-news.britain.eu.net!lyra.csx.cam.ac.uk!dh124 From: dh124@cus.cam.ac.uk (D. Hodges) Newsgroups: bionet.molbio.proteins Subject: Re: glycosylation Date: 6 Dec 1995 08:26:41 GMT Organization: University of Cambridge, England Lines: 20 Message-ID: <4a3k41$g6s@lyra.csx.cam.ac.uk> References: <4a15tm$krn@saphir.jouy.inra.fr> NNTP-Posting-Host: grus.cus.cam.ac.uk X-Newsreader: TIN [version 1.2 PL2] François Villatte (villatte@versailles.inra.fr) wrote: : Hello, : I currently search a colorimetric method to detect glycoproteins on a : native PAGE. : Moreover, I would like to know if a GST detection method on PAGE exist. : Francois For detecting glycoproteins try probing a western blot of your gel with biotinylated lectins of various types to cover the different binding affinities. Then use streptavidin conjugated hores-radish peroxidase as the second step and finally develop the reaction using any colour generating HRP substrate. I have used 4-chloronapthol quite successfully. Debbie -- --------------------------------------------------------------------------- Debbie Hodges dh124@cus.cam.ac.uk Fax 01223 217838 Rheumatology Research Unit, Addenbrookes Hospital, Hills Road, Cambridge, UK. ---------------------------------------------------------------------------- I never could get the hang of Thursdays (A.Dent). From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!news.ksu.ksu.edu!usenet From: "Dr. Laura A. Andersson" Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: 6 Dec 1995 17:42:25 GMT Organization: Kansas State University Lines: 3 Message-ID: <4a4km1$ctv@newserv.ksu.ksu.edu> References: NNTP-Posting-Host: 129.130.71.12 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) To: Bob_Hoesch@fws.gov Xref: biosci bionet.general:18778 bionet.molbio.proteins:6431 bionet.population-bio:1696 sci.chem:44487 Hi. the heme protein literature is rife with out-dated/poorly defined terminology. In this case, "reduction" of methemoglobin DOES NOT mean reduction in terms of From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!news.ksu.ksu.edu!usenet From: "Dr. Laura A. Andersson" Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: 6 Dec 1995 17:40:20 GMT Organization: Kansas State University Lines: 2 Message-ID: <4a4ki4$ctv@newserv.ksu.ksu.edu> References: NNTP-Posting-Host: 129.130.71.12 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) To: Bob,Hoesch Xref: biosci bionet.general:18777 bionet.molbio.proteins:6430 bionet.population-bio:1695 sci.chem:44486 Hi. From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!EU.net!howland.reston.ans.net!newsfeed.internetmci.com!uwm.edu!msunews!netnews.upenn.edu!cronkite.ocis.temple.edu!astro.ocis.temple.edu!kevo From: kevo@astro.ocis.temple.edu (KevO) Newsgroups: bionet.molbio.proteins Subject: Re: Disappearing SDS-PAGE bands Date: 7 Dec 1995 05:52:58 GMT Organization: Temple University, Academic Computer Services Lines: 29 Message-ID: <4a5vfq$nsg@cronkite.ocis.temple.edu> References: <30C36A66.564B@oci.utoronto.ca> NNTP-Posting-Host: astro.ocis.temple.edu X-Newsreader: TIN [version 1.2 PL2] Kelly Ambler (kelly@hearts.bsd.uchicago.edu) wrote in article : :>I don't think it was the staining conditions - I ran, and thus stained, :>different individual gels (all polymerized in one batch) on separate [some stuff deleted] :>These were 11% acrylamide gels. :>One other fact - the demarcation between bands and no-bands was quite :>sharp at the bottom of the "problem zone", but appeared to fade out :>gradually at the top of the "problem zone". This is a definate puzzler. From what I hear, I have a strong feeling that the gel did not polymerize well. (Incomplete crosslinking). This could explain the the bands are "clearer" at the bottom (I am assuming that you are using a vertical slab gel). I would suggest that you remake your monomer solution using FRESH material. That should do the trick. Also, one of my fellow student here (actually brought this up a while ago) claims that we had to leave our gel to polymerize at least 4-5 hours (we run BioRad minigels) in order for proper polymerization. Let me know how it goes. Good luck! -- ****************************************************************************** * Kevin Ong * For God so loved the world that he * * Biology Dept. * gave his only begotten son... * * Temple University,Philadelphia * -JOHN 3:16 * * O:(215)204-8843 H:(610)352-4773 *************************************** * Fax:(215)204-6646 email> kevo@astro.ocis.temple.edu or kevo@vm.temple.edu * * WWW URL> http://astro.temple.edu/~kevo * ****************************************************************************** From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!chi-news.cic.net!uwm.edu!msunews!netnews.upenn.edu!cronkite.ocis.temple.edu!astro.ocis.temple.edu!driska From: driska@astro.ocis.temple.edu (Stephen P. Driska PhD) Newsgroups: bionet.molbio.proteins Subject: ATP-gamma-S and tyrosine kinases Date: 6 Dec 1995 23:06:07 GMT Organization: Temple University, Academic Computer Services Lines: 18 Message-ID: <4a57kv$dva@cronkite.ocis.temple.edu> NNTP-Posting-Host: astro.ocis.temple.edu Summary: Does anyone know if ATP-gamma-S is a substrate for tyrosine kinases? Keywords: Locks, combinations, doorknobs, bolts, deadbolts, master, major, and minor. X-Newsreader: TIN [version 1.2 PL2] The compound ATP-gamma-S has been used for twenty years because it is a substrate for many serine/threonine kinases, but the thiophosphate group incorporated into the protein is (relatively) resistant to phosphoprotein phosphatases. Thus, it provides a way to essentially permanently "turn on" a system which is normally only transiently activated. I was wondering if this compound was a substrate for tyrosine kinases, and if so, whether any such thiophosphorylated (on tyrosine) proteins would be resistant to the action of phosphotyrosine phosphatases. Just wondering. Thanks for any information -- Steve Driska, Physiology Department, Temple University Medical School Philadelphia, PA 19140 USA (215) 707-3283 driska@astro.ocis.temple.edu If I could think of something witty and clever to say, it would go right here ----->> ......................... From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!vixen.cso.uiuc.edu!news.ksu.ksu.edu!usenet From: "Dr. Laura A. Andersson" Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: 6 Dec 1995 17:55:15 GMT Organization: Kansas State University Lines: 9 Message-ID: <4a4le3$d1n@newserv.ksu.ksu.edu> References: NNTP-Posting-Host: 129.130.71.12 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) To: Bob_Hoesch@fws.gov Xref: biosci bionet.general:18779 bionet.molbio.proteins:6432 bionet.population-bio:1697 sci.chem:44489 trying again. [this is my first time on newsgroup]. anyway, the "reduction" is not a redox reaction, but a "decrease" (also a reduction) in amount of metMb. With respect to number of hemoglobin bands, first metHb has bands at 405 (Soret) 500 & 629 nm, with millimolar extinction coefficients, respectively, of 179, 10, and 4.4. but for cyanometHb, the bands are at 418 and 541 nm, with millimolar extinction coefficeints of 124 and 12.5 nm. key point - the "high-spin" marker at 629 is GONE if the cyanide form is complete. [Hb has a high affinity for cyanide - only need ~10 x to be complete.] From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!fdn.fr!jussieu.fr!univ-lille1.fr!cresimm-pc1.univ-lille1.fr!vergoten From: vergoten@pop.univ-lille1.fr (Gerard Vergoten) Newsgroups: bionet.molbio.proteins Subject: NATO-ASI on Biomolecular structure and dynamics"NATO Advanced Study Institute" Date: Wed, 6 Dec 1995 17:50:57 Organization: CRESIMM - C8 - USTL - Villeneuve d'Ascq Lines: 15 Distribution: world Message-ID: NNTP-Posting-Host: cresimm-pc1.univ-lille1.fr Keywords: molecular modeling, computer simulations X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] "BIOMOLECULAR STRUCTURE AND DYNAMICS: RECENT EXPERIMENTAL AND THEORETICAL ADVANCES" ASI LOUTRAKI, Greece May 27-June 6,1996 Contact: Professor G. VERGOTEN Address: Université des Sciences et Technologies de Lille CRESIMM ( U 279 INSERM ) UFR de Chimie Bât C8 - 1er étage 59655 VILLENEUVE D'ASCQ FRANCE FAX : (33) 20 33 72 79 E mail : vergoten@pop.univ-lille1.fr Designated Publisher: KLUWER From owner-proteins@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!EU.net!peer-news.britain.eu.net!zippy.dct.ac.uk!dundee.ac.uk!dundee.ac.uk!not-for-mail Newsgroups: bionet.molbio.proteins Subject: Re: glyco Message-ID: <4a4gho$891@dux.dundee.ac.uk> From: hjtreuma@dux.dundee.ac.uk (H.J. Treumann Biochemistry ext 4301) Date: 6 Dec 1995 16:31:52 -0000 References: <4a1qr1$247@saphir.jouy.inra.fr> <4a4325$cno@dux.dundee.ac.uk> Organization: The University, Dundee, DD1 4HN, Scotland, UK. NNTP-Posting-Host: dux.dundee.ac.uk X-Newsreader: TIN [version 1.2 PL1] Lines: 21 Hello, sorry about the previous reply - tin ate it :-( I suggested the use of stains-all (ref: Green et al. (1973), Anal. Biochem. Dahlberg et al. (1969), J. Mol. Biol. 41, pp. 139-147). Alternatively you could oxidise your gels with periodic acid before you silver stain them. Both methods are not selective for glycoproteins but they might help you to detect proteins that remain invisible with the standard staining methods. Good luck, Achim -- Achim Treumann tel. +44-1382-344301 Carbohydrate Research Centre fax +44-1382-322583 Department of Biochemistry Dundee DD1 4HN, Scotland, UK email a.treumann@dundee.ac.uk From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!daresbury!not-for-mail From: "Andrew, Tel. +396-91093434" Newsgroups: bionet.molbio.proteins Subject: RE: biotinylation Date: 7 Dec 1995 15:55:55 -0000 Lines: 42 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <4a72qb$i2j@mserv1.dl.ac.uk> Original-To: proteins@dl.ac.uk [apologies for replying here, mail to the original sender's address bounces] Hello Mara, In bionet.molbio.proteins Msg # 5991 you wrote: >I'm currently trying to biotinylate a protein that is known to have 3 free >cysteine residues (not involved in disulfide bridges) using iodoacetyl-biotin >to specifically modify the sulfhydryl groups. But for some reason it's not >working. The protocol that comes with the reagent isn't quite appropriate for >what I'm doing, so it's not very helpful. Another possible problem is that the >protein sample I'm working with was aggregated. I dis-aggregated it with SDS, >and it looked okay on a gel, but still... >Has anyone used this reagent? Can you offer me any advice? I've never used that particular reagent so I can't help you with it directly, but my experience of iodoacetamides is that they are not terribly reactive with sulfhydryl groups. If possible, I would try to change to something like a maleimido-biotin derivative (e.g. Biotin-BMCC) since this should be more reactive. You can buy the stuff from Pierce (catalog # 21900) Pierce Tel. 800-874-3723 or 815-968-0747. Note that I don't work for Pierce, I'm just a satisfied customer. Andrew |========================================================================| | Andrew Wallace | Discussion on phage display, | | | combinatorial libraries, etc. - | | IRBM P. Angeletti, | bionet.molecules.repertoires | | Via Pontina KM 30.600 | molreps@daresbury.ac.uk | | 00040 Pomezia, Italy. |-------------------------------------------| | | "It has not escaped our notice that | | Voice: +39-6-91093434 | the specific pairing we have postulated | | Fax: +39-6-91093225 | immediately suggests a possible copying | | Email: wallace@irbm.it | mechanism for the genetic material." | | | | | DISCLAIMER: I do not speak | J.D. Watson and F.H.C. Crick | | on behalf of anyone. | in Nature 171:737-737 (1953). | |========================================================================| From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Path: biosci!agate!howland.reston.ans.net!newsfeed.internetmci.com!news.kei.com!newsstand.cit.cornell.edu!newsfeed.cit.cornell.edu!cornellcs!rochester!galileo.cc.rochester.edu!uhura.cc.rochester.edu!gl002c From: gl002c@uhura.cc.rochester.edu (G. Lane) Subject: Re: Hemoglobin and Cyanide Message-ID: <1995Dec7.060924.29134@galileo.cc.rochester.edu> Sender: news@galileo.cc.rochester.edu Nntp-Posting-Host: uhura.cc.rochester.edu Organization: University of Rochester - Rochester, New York References: <4a4le3$d1n@newserv.ksu.ksu.edu> Date: Thu, 7 Dec 95 06:09:24 GMT Lines: 15 Xref: biosci bionet.general:18795 bionet.molbio.proteins:6436 bionet.population-bio:1698 sci.chem:44526 In <4a4le3$d1n@newserv.ksu.ksu.edu> "Dr. Laura A. Andersson" writes: >trying again. [this is my first time on newsgroup]. anyway, the "reduction" is not >a redox reaction, but a "decrease" (also a reduction) in amount of metMb. With >respect to number of hemoglobin bands, first metHb has bands at 405 (Soret) 500 & >629 nm, with millimolar extinction coefficients, respectively, of 179, 10, and 4.4. > but for cyanometHb, the bands are at 418 and 541 nm, with millimolar extinction >coefficeints of 124 and 12.5 nm. key point - the "high-spin" marker at 629 is >GONE if the cyanide form is complete. [Hb has a high affinity for cyanide - only >need ~10 x to be complete.] is the cyanide strength with Hb at the iron, leading one to a similar conclusion with carbon monoxide (oxygen is worse at bonding to iron than either CN- , or CO), My question: does exposure to cyanide resemble carbon monoxide exposure? From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!news.cac.psu.edu!news.math.psu.edu!hudson.lm.com!godot.cc.duq.edu!newsfeed.pitt.edu!dsinc!ub!acsu.buffalo.edu!ubvms.cc.buffalo.edu!v059lr6y From: v059lr6y@ubvms.cc.buffalo.edu (Timothy E Berlinski) Newsgroups: bionet.molbio.proteins Subject: Extraction help? Date: 7 Dec 1995 07:43 EST Organization: University at Buffalo Lines: 12 Distribution: world Message-ID: <7DEC199507430124@ubvms.cc.buffalo.edu> NNTP-Posting-Host: ubvmsb.cc.buffalo.edu News-Software: VAX/VMS VNEWS 1.41 I wrote before and maybe I confuse my situation a bit. I have a hypothesis I wish to persue. And I think that it has some substance to it. But the problem is that I don't know how to go about doing it. See I need to know how to seperate and purify DNA, and RNA from cell extracts. I know that there is an ethanol precipitation method to DNA extraction but you just don't go and add 10 ml of ethanol to your sample and hope your DNA precipitates, you see what I need now? I can get the resources to do this but I need to know what to use, how to use it and how much to use. Thank You. Tim Berlinski From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!torn!news.bc.net!rover.ucs.ualberta.ca!tribune.usask.ca!canopus.cc.umanitoba.ca!cc-hsc-k-box26.cc.umanitoba.ca!user From: egreif@cc.umanitoba.ca (Elke) Newsgroups: bionet.molbio.proteins Subject: Re: Disappearing SDS-PAGE bands Date: Thu, 07 Dec 1995 12:24:32 -0600 Organization: University of Manitoba Lines: 38 Message-ID: References: <30C36A66.564B@oci.utoronto.ca> NNTP-Posting-Host: cc-hsc-k-box26.cc.umanitoba.ca In article , Kelly Ambler wrote: > I don't think it was the staining conditions - I ran, and thus stained, > different individual gels (all polymerized in one batch) on separate > days, yet had the same band pattern in all the gels. I reuse my > staining solution and did not have this problem with other gels stained > at later date. > > These were 11% acrylamide gels. > > One other fact - the demarcation between bands and no-bands was quite > sharp at the bottom of the "problem zone", but appeared to fade out > gradually at the top of the "problem zone". > > I have talked to several people about this problem, and we're all quite > puzzled. Mostly, I would like to ensure that it doesn't occur again! > > Kelly Ambler > University of Chicago Hi Kelly, I've been running SDS-PAGE for years and have also come across your problem. I switched to 7.5%-15% gradient gels and the bands were tight and all were visible as compared to before. Also, I increased the staining time with Coomassie from 45 mins to 1.5 hours. It seemed to help to. Take a look at the gel while it's staining and make sure the entire gel is covered, I know it sounds ridiculous but stranger things have happened, by the way this is science, right? Anyway, if you try gradients, let me know, we could also use the feedback. Good luck!!! Elke -- Hey, hey, hey, hey! It was the DNA. Hey, hey, hey, hey! That made me this way. From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!rutgers!csn!carbon!night.primate.wisc.edu!sdd.hp.com!swrinde!howland.reston.ans.net!nntp.coast.net!zombie.ncsc.mil!cs.umd.edu!haven.umd.edu!news.ums.edu!umabnet.ab.umd.edu!umabnet.ab.umd.edu!mremingt From: "Mary P. Remington" Newsgroups: bionet.molbio.proteins Subject: SAAB protein program Date: Thu, 7 Dec 1995 19:27:53 -0500 Organization: University of Maryland at Baltimore Lines: 4 Message-ID: NNTP-Posting-Host: umabnet.ab.umd.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Is anyone familiar with a program called SAAB which apparently searches for and identifies protein consensus sequences which are protein binding sites. Any input appreciated. Thanks, Mary From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!newsfeed.pitt.edu!godot.cc.duq.edu!news.duke.edu!usenet From: Mara Michelle Casar Newsgroups: bionet.molbio.proteins Subject: biotinylation Date: 7 Dec 1995 13:48:17 GMT Organization: Duke University, Durham, NC, USA Lines: 19 Message-ID: <4a6rb1$17l@news.duke.edu> NNTP-Posting-Host: north8.acpub.duke.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (X11; I; SunOS 5.3 sun4m) X-URL: news:bionet.molbio.proteins Hi - I'm a first-year grad student at Duke and brand-new to this group. I have a problem that I hope someone out there can help me with... I'm currently trying to biotinylate a protein that is known to have 3 free cysteine residues (not involved in disulfide bridges) using iodoacetyl-biotin to specifically modify the sulfhydryl groups. But for some reason it's not working. The protocol that comes with the reagent isn't quite appropriate for what I'm doing, so it's not very helpful. Another possible problem is that the protein sample I'm working with was aggregated. I dis-aggregated it with SDS, and it looked okay on a gel, but still... Has anyone used this reagent? Can you offer me any advice? Thanks in advance, Mara From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!geraldo.cc.utexas.edu!bashful.cc.utexas.edu!hantash From: hantash Newsgroups: bionet.molbio.proteins Subject: Magic Bands Date: Thu, 7 Dec 1995 17:14:42 -0600 Organization: The University of Texas at Austin, Austin, Texas Lines: 16 Message-ID: NNTP-Posting-Host: bashful.cc.utexas.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Hi All; I perform western blots on some bacterial proteins. For some of the proteins When I compare total cells fraction to periplasmic fraction, a band that is not present in total cells shows up in the periplasmic fraction of the same cell !! Usually one should see the band in both total cells and periplasmic fraction. I tried to reason it by assuming that the band that shows up in the periplasmic fraction is masked in the lane for total cells fraction by other proteins. Any other ideas or comments or suggestions ? Feras hantash@ccwf.cc.utexas.edu From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!howland.reston.ans.net!math.ohio-state.edu!magnus.acs.ohio-state.edu!csn!borcore.usbr.gov!sacsa3.mp.usbr.gov!fwssun1.irm.r6.fws.gov!ash.lab.r1.fws.gov!bob!Bob_Hoesch From: Bob_Hoesch@fws.gov (Bob Hoesch) Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: Thu, 7 Dec 1995 12:33:58 Organization: U.S. Fish & Wildlife Service Forensics Laboratory Lines: 19 Message-ID: References: <4a4le3$d1n@newserv.ksu.ksu.edu> NNTP-Posting-Host: bob.lab.r1.fws.gov X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Xref: biosci bionet.general:18805 bionet.molbio.proteins:6443 bionet.population-bio:1699 sci.chem:44567 In article <4a4le3$d1n@newserv.ksu.ksu.edu> "Dr. Laura A. Andersson" writes: >From: "Dr. Laura A. Andersson" >Subject: Re: Hemoglobin and Cyanide >Date: 6 Dec 1995 17:55:15 GMT > the "reduction" is not >a redox reaction, but a "decrease" (also a reduction) in amount of metMb. With >respect to number of hemoglobin bands, first metHb has bands at 405 (Soret) 500 & >629 nm, with millimolar extinction coefficients, respectively, of 179, 10, and 4.4. > but for cyanometHb, the bands are at 418 and 541 nm, with millimolar extinction >coefficeints of 124 and 12.5 nm. key point - the "high-spin" marker at 629 is >GONE if the cyanide form is complete. [Hb has a high affinity for cyanide - only >need ~10 x to be complete.] This makes sense in terms of a change in the number of spectrophotometric absorption peaks, but I still don't think we have a good explanation for the reduction in the number of electrophoretic bands. After all, the protein doesn't go away. From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!howland.reston.ans.net!vixen.cso.uiuc.edu!news.ecn.bgu.edu!psuvax1!news.eecs.nwu.edu!newsfeed.acns.nwu.edu!news.cc.uic.edu!usenet From: Keld Sorensen Newsgroups: bionet.molbio.proteins Subject: Re: biotinylation Date: 7 Dec 1995 20:45:53 GMT Organization: Univ. Illinois / College of Med. Lines: 7 Message-ID: <4a7jq1$2nvq@tigger.cc.uic.edu> References: <4a6rb1$17l@news.duke.edu> NNTP-Posting-Host: sliprock-03.dialin.uic.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; PPC) X-URL: news:4a6rb1$17l@news.duke.edu Are the sulfhydryls really available, i.e. have you done an Ellman assay?? Also, you may want to try Biotin-BMCC (similar to the reagent you are using, but with better characteristics) Keld. From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!rutgers!csn!magnus.acs.ohio-state.edu!lerc.nasa.gov!purdue!oitnews.harvard.edu!news.dfci.harvard.edu!usenet From: "Matthew P. Frosch, M.D., Ph.D." Newsgroups: bionet.molbio.proteins Subject: Re: Ligase efficiency in different buffers ? Date: 7 Dec 1995 21:47:32 GMT Organization: Brigham and Women's Hospital Lines: 4 Message-ID: <4a7ndk$h9n@cisunix1.dfci.harvard.edu> References: NNTP-Posting-Host: 134.174.88.158 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) X-URL: news:stefanba-0712951939580001@mun1.mun.uu.se TRY CONTACTING THE FOLKS AT NEB (http://www.neb.com/) THEY ARE VERY HELPFUL ABOUT TRICKS FOR USING THEIR ENZYMES From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!uwm.edu!post.its.mcw.edu!usenet From: Jennifer Potter Newsgroups: bionet.molbio.proteins Subject: Re: Hela Cell 2-d map? Date: 7 Dec 1995 22:34:13 GMT Organization: Medical College of Wisconsin Lines: 15 Message-ID: <4a7q55$jmv@post.its.mcw.edu> References: NNTP-Posting-Host: d350-1.biochem.mcw.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) To: mdeziel@albany.net Dear Dr. Deziel, I have a reference for you regarding a 2-D HeLa Cell map. Unfortunately on my copy the journal name, etc. was not copied (sorry!). What I can give you is the authors - Donald J. Higgens and Thomas W. Conway. The paper I have was published in 1988 where these authors looked at IFN induced proteins in Hela cells. Sorry I don't have anything more for you, but at least this may point you in the right direction. Best of luck, Jennifer L. Potter Medical College of WI From owner-proteins@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!howland.reston.ans.net!nntp.coast.net!news00.sunet.se!sunic!news99.sunet.se!columba.udac.uu.se!mun1.mun.uu.se!user From: stefanba@bio.embnet.se (Stefan Bäckström) Newsgroups: bionet.molbio.proteins Subject: Ligase efficiency in different buffers ? Date: Thu, 07 Dec 1995 19:39:58 +0100 Organization: Department of Developmental Neuroscience Lines: 9 Message-ID: NNTP-Posting-Host: mun1.mun.uu.se Hello. Does anyone know of a review on how well T4 DNA ligase works in different buffers ? Specially restriction buffers ? Thanks, Stefan Bäckström From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!ihnp4.ucsd.edu!agate!howland.reston.ans.net!newsfeed.internetmci.com!in2.uu.net!brighton.openmarket.com!decwrl!lll-winken.llnl.gov!fnnews.fnal.gov!nntp-server.caltech.edu!kaihsu From: kaihsu@whip.ugcs.caltech.edu (Kai-hsu TAI) Newsgroups: bionet.molbio.proteins Subject: [help] protein design Date: 8 Dec 1995 23:49:45 GMT Organization: California Institute of Technology, Pasadena Lines: 10 Message-ID: <4aaiup$co2@gap.cco.caltech.edu> NNTP-Posting-Host: whip.ugcs.caltech.edu X-Newsreader: TIN [version 1.2 PL2] I am a college sophomore majoring chemistry. Recently I start to discover my interest in protein design. Would anyone please tell me how to get to know more about this field, e.g., what kind of courses to take, what books to read. Thank you very much for your attention. Cheers, Kai-hsu Tai Soph. Chem. Caltech http://www.ugcs.caltech.edu/~kaihsu/ kaihsu@ugcs.caltech.edu From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!ns1.faseb.org!lamarck.sura.net!ra.nrl.navy.mil!news.math.psu.edu!psuvax1!gvls1!newsfeed.pitt.edu!dsinc!spool.mu.edu!howland.reston.ans.net!nntp.coast.net!torn!watserv3.uwaterloo.ca!rustfly From: ACSCOTT@SCIENCE.uwaterloo.ca (Andrew C. Scott) Subject: Help looking for uses of Snake Venoms and components Message-ID: <4a9jh7$8ho_001@watstar.uwaterloo.ca> Sender: news@watserv3.uwaterloo.ca Nntp-Posting-Host: rustfly.watstar.uwaterloo.ca Organization: University of Waterloo X-Newsreader: News Xpress Version 1.0 Beta #3 Date: Fri, 8 Dec 1995 14:53:26 GMT Lines: 20 Hello... I am doing some market research for developing a company which produces snake venoms and possibly the purified components (peptides) of the venoms. I was wondering if anyone could tell me what uses such venoms have out there and if there is much of a demand for them. Also, if I could be directed to anyone who uses them on a regular basis for research I'd appreciate it. If anyone has any information at all, I'd appreciate it and thank you for your time. Sincerely, Andrew C. Scott ------------------------------------------------------------------------------ Andrew C. Scott email: acscott@sciborg.uwaterloo.ca VENOMTECH Inc. acscott@science.watstar.uwaterloo.ca ------------------------------------------------------------------------------ From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!daresbury!nntp-trd.UNINETT.no!newsfeed.sunet.se!news00.sunet.se!sunic!news99.sunet.se!news.funet.fi!news.eunet.fi!newsmaster From: piero pollesello Newsgroups: bionet.molbio.proteins Subject: Re: Magic Bands Date: 8 Dec 1995 09:41:36 GMT Organization: POLYbios, Area Science Park, Trieste, Italy Lines: 13 Message-ID: <4a918g$6os@idefix.eunet.fi> References: NNTP-Posting-Host: mac545.espoo.orion.fi Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; 68K) X-URL: news:Pine.SUN.3.91.951207170906.6844B-100000@bashful.cc.utexas.edu (1) a band could appear due to dimerization (-S-S-, do you run in presence of Mercaptoethanol?) or detergent resistant clustering (phospholamban is pentamerizing even if you boil it with SDS) (2) a band could appear due to degradation (some protease are present? do you obtain your fractions in presence of protease inhibitors? Which?) You gave too few informations for a reliable diagnosys of your problem, if it is a problem. Piero From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!ns1.faseb.org!lamarck.sura.net!ra.nrl.navy.mil!news.math.psu.edu!psuvax1!news.eecs.nwu.edu!newsfeed.acns.nwu.edu!news.luc.edu!chi-news.cic.net!nntp.coast.net!torn!utnut!oci!news From: Flip Hoedemaeker Subject: Re: Protein Models & CAD/CAM Content-Type: text/plain; charset=us-ascii Message-ID: <30C8932B.5862@oci.utoronto.ca> Sender: news@oci.utoronto.ca Content-Transfer-Encoding: 7bit Organization: The Holistic Detective Agency References: <4a2rre$alp@agate.berkeley.edu> Mime-Version: 1.0 Date: Fri, 8 Dec 1995 19:34:03 GMT X-Mailer: Mozilla 2.0b2a (Windows; I; 16bit) Lines: 14 Louis Hom wrote: > > It seems to me that life for some folks would be easier if you could have > your computer create a real 3D representation of a protein structure. I > know that computer-controlled generation of prototypes in manufacturing is > done all the time. Does anyone know of someone trying to apply computer > aided manufacturing technology to protein structures? I'm not sure what you mean with "manufacturing technology" , but the SGI's we use to model protein structures obtained by X-ray and NMR are exactly the same as what car or plane or computer animation designers use. Of course the software is different, but we get to see "real" 3D images! Flip From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!FOX.NSTN.CA!ewright From: ewright@FOX.NSTN.CA ("Edwin Wright") Newsgroups: bionet.molbio.proteins Subject: Book Entitled, "Cytochromes c" Date: 8 Dec 1995 16:42:26 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: <84713.ewright@fox.nstn.ca> NNTP-Posting-Host: net.bio.net Does anyone have an email address for Geoffrey R. Moore or Graham W. Pettigrew, co-authors of the subject book? Regards, -- Edwin Wright ewright@fox.nstn.ca From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!newshost.lanl.gov!ncar!uchinews!vixen.cso.uiuc.edu!newsfeed.internetmci.com!in2.uu.net!news.mtholyoke.edu!news From: "Sean M. Decatur" Newsgroups: bionet.general,bionet.molbio.proteins,bionet.population-bio,sci.chem Subject: Re: Hemoglobin and Cyanide Date: 8 Dec 1995 21:23:52 GMT Organization: Mount Holyoke College Lines: 19 Message-ID: <4aaad8$4d6@mudraker.mtholyoke.edu> References: <4a4le3$d1n@newserv.ksu.ksu.edu> NNTP-Posting-Host: 138.110.18.26 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: quoted-printable X-Mailer: Mozilla 1.1N (Macintosh; I; PPC) To: Bob_Hoesch@fws.gov X-URL: news:Bob_Hoesch.453.000C9153@fws.gov Xref: biosci bionet.general:18826 bionet.molbio.proteins:6455 bionet.population-bio:1702 sci.chem:44615 Bob_Hoesch@fws.gov (Bob Hoesch) wrote: >This makes sense in terms of a change in the number of spectrophotometric >absorption peaks, but I still don't think we have a good explanation for the >reduction in the number of electrophoretic bands. After all, the protein >doesn't go away. > Heres a thought-- When CN- binds to metHb, the net charge of the protein is the same as deoxy- or oxyHb, whereas metHb (with H2O bound) has net charge= of +1 relative to the reduced (oxy- and dexoy-) species. Thus, if there is a mixture of reduced and oxidized irons present in the = Hb solution, there will be a mixture of pIs (and presumably multiple IEF bands). However, when CN- is added the metHbCN will have t= he same net charge as the oxyHb and/or deoxyHb, so the Hbs should all have the same pI. From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!CS.Arizona.EDU!news.Arizona.EDU!hamblin.math.byu.edu!acs2.byu.edu!news.cuny.edu!news.sprintlink.net!rain.fr!jussieu.fr!univ-lyon1.fr!in2p3.fr!oleane!plug.news.pipex.net!pipex!tube.news.pipex.net!pipex!tank.news.pipex.net!pipex!newsfeed.internetmci.com!howland.reston.ans.net!newsserver.jvnc.net!raffles.technet.sg!nova.np.ac.sg!nova.np.ac.sg!news From: 93462036@comet.np.ac.sg (Ang Ching Seng) Newsgroups: bionet.molbio.proteins Subject: Help !!! Reduction of Hemocyanin(KLH) Date: 7 Dec 1995 12:56:16 +0800 Organization: Ngee Ann Polytechnic, Singapore Lines: 13 Message-ID: <4a5s5g$471@comet.np.ac.sg> NNTP-Posting-Host: comet.np.ac.sg NNTP-Posting-User: 93462036 Dear people, Can anyone tell me how can i reduce KLH ( Hemocyanin from keyhole Limpets) so as to expose its thiol (-SH) groups ? I need information such as the buffers to dissolve it in as i've tried a few buffers but the protein doesn't seem to dissolve. Appreciate very much if anyone would reply. THANKS !!!!! Ang Ching Seng 93462036@np.ac.sg From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!biosci!not-for-mail From: biohelp@net.bio.net (BIOSCI Administrator) Newsgroups: bionet.molbio.methds-reagnts,bionet.general,bionet.genome.arabidopsis,bionet.software,bionet.molbio.proteins,bionet.neuroscience Subject: IMPORTANT! - net.bio.net down for backups Sunday Date: 8 Dec 1995 11:42:06 -0800 Organization: BIOSCI International Newsgroups for Biology Lines: 13 Distribution: world Message-ID: <4aa4ee$9kt@net.bio.net> NNTP-Posting-Host: net.bio.net Xref: biosci bionet.molbio.methds-reagnts:37372 bionet.general:18817 bionet.genome.arabidopsis:4023 bionet.software:14124 bionet.molbio.proteins:6452 bionet.neuroscience:11601 NOTE!! The U.S. BIOSCI node at net.bio.net will be down this Sunday from 10 AM Pacific Time to about 6 PM for backups. Mail sent to the newsgroups and USENET articles should remain queued during this time, but access to WWW, gopher and WAIS services will be unavailable. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!CS.Arizona.EDU!news.Arizona.EDU!hamblin.math.byu.edu!acs2.byu.edu!news.cuny.edu!news.sprintlink.net!newsfeed.internetmci.com!howland.reston.ans.net!usc!news.cerf.net!newsserver.sdsc.edu!news.tc.cornell.edu!newsstand.cit.cornell.edu!NewsWatcher!user From: mcb10@cornell.edu (Peggy Barr) Newsgroups: bionet.molbio.hiv,bionet.molbio.proteins Subject: Re: 70 kd non-viral banding in HIV WB Date: Fri, 08 Dec 1995 10:08:59 -0400 Organization: Cornell University Lines: 25 Sender: mcb10@cornell.edu (Verified) Message-ID: References: <4a9avm$skc$1@mhade.production.compuserve.com> NNTP-Posting-Host: 128 Xref: biosci bionet.molbio.hiv:1776 bionet.molbio.proteins:6458 In article <4a9avm$skc$1@mhade.production.compuserve.com>, Jonathan Trouern-Trend <75551.567@CompuServe.COM> wrote: > I'm doing some work on HIV indeterminate banding patterns in > Western Blots and found that over 20% of IWB (Indet. Western Blot) > have a non-viral band at 70kd. There has been alot of literature > about IWBs in general, like those with only the GAG protein bands > but I haven't come across any explanations for these bands. > Perhaps they are considered uninteresting artifacts. Any info > would be appreciated. > Jonathan Trouern-Trend > Univ. of CT ************************************************************************** I do a lot of Western blotting for detection of antibodies to feline immunodeficiency virus. The primary non-viral bands in these blots are also in the 60 to 70 kD range and appear to be due to reactivity with albumin and other cell culture components. We assume that much of the reactivity is due to previous vaccination of the cats with killed or MLV products grown in tissue culture systems. Peggy Barr College of Vet. Med. Cornell Univ. From owner-proteins@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!CS.Arizona.EDU!news.Arizona.EDU!hamblin.math.byu.edu!sol.ctr.columbia.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!EU.net!peer-news.britain.eu.net!zippy.dct.ac.uk!dundee.ac.uk!dundee.ac.uk!not-for-mail Newsgroups: bionet.molbio.proteins Subject: PO4 - detergent precipitate/hydroxyapatite Message-ID: <4aa0vk$iao@dux.dundee.ac.uk> From: hjtreuma@dux.dundee.ac.uk (H.J. Treumann Biochemistry ext 4301) Date: 8 Dec 1995 18:43:00 -0000 Organization: The University, Dundee, DD1 4HN, Scotland, UK. NNTP-Posting-Host: dux.dundee.ac.uk X-Newsreader: TIN [version 1.2 PL1] Lines: 24 Dear colleagues, we are trying to purify a proand want to use a hydroxyapatite column for that purpose. The chromatography has to occur at 4 C for obvious reasons. The protein we are dealing with is a membrane protein which is soluble only in the presence of detergent. As a detergent we are using N-octyl-glucoside (NOG). We have encountered the following problem: we get a precipitate of the detergent in phosphate buffers in the HPLC pump (in the bottle the buffers seem to be perfectly soluble). Has anyone else experienced these difficulties? We would very much appreciate input from anybody (in the newsgroup or by email). If there is sufficient interest I will post a summary on the net. Thank you very much in advance, Achim -- Achim Treumann tel. +44-1382-344301 Carbohydrate Research Centre fax +44-1382-322583 Department of Biochemistry Dundee DD1 4HN, Scotland, UK email a.treumann@dundee.ac.uk From owner-proteins@net.bio.net Fri Dec 08 22:00:00 1995 Path: biosci!ihnp4.ucsd.edu!agate!howland.reston.ans.net!newsfeed.internetmci.com!chi-news.cic.net!nntp.coast.net!news00.sunet.se!sunic!news99.sunet.se!news.rccn.net!gnu.mat.uc.pt!histocentro From: psantos@gemini.ci.uc.pt (Paulo Santos) Newsgroups: bionet.molbio.proteins Subject: Biosensors (MHC-peptide-TCR) Date: 9 Dec 1995 18:18:11 GMT Organization: LUSOTRANSPLANTE Lines: 25 Message-ID: <4acjt3$4vr@gnu.mat.uc.pt> NNTP-Posting-Host: histocentro.uc.pt X-Newsreader: News Xpress Version 1.0 Beta #3 I would like to have some information about biosensors and it's use for analysing molecules interaction. My special interest is the aplication to the MHC-peptide and TCR affinities (study the interaction of a special MHC allele and a pool of naturally or synthetic peptides, or vice-versa, a special peptide and a pool of MHC alleles). Thanks in advance. Paulo Santos Laboratorio de Genetica Molecular Centro de Histocompatibilidade do Centro LUSOTRANSPLANTE Faculdade de Medicina da Universidade de Coimbra, piso 3 Apartado 3004 P-3049 COIMBRA Codex PORTUGAL Telefone +351-39-33693 +351-39-20527 Fax +351-39-33674 Email: psantos@gemini.ci.uc.pt From owner-proteins@net.bio.net Fri Dec 08 22:00:00 1995 Path: biosci!ihnp4.ucsd.edu!swrinde!newsfeed.internetmci.com!news.compuserve.com!news.production.compuserve.com!news From: Jonathan Trouern-Trend <75551.567@CompuServe.COM> Newsgroups: bionet.molbio.hiv,bionet.molbio.proteins Subject: 70 kd non-viral banding in HIV WB Date: 8 Dec 1995 12:27:34 GMT Organization: CompuServe, Inc. (1-800-689-0736) Lines: 9 Message-ID: <4a9avm$skc$1@mhade.production.compuserve.com> Xref: biosci bionet.molbio.hiv:1778 bionet.molbio.proteins:6460 I'm doing some work on HIV indeterminate banding patterns in Western Blots and found that over 20% of IWB (Indet. Western Blot) have a non-viral band at 70kd. There has been alot of literature about IWBs in general, like those with only the GAG protein bands but I haven't come across any explanations for these bands. Perhaps they are considered uninteresting artifacts. Any info would be appreciated. Jonathan Trouern-Trend Univ. of CT From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins Subject: *IMPORTANT* New BIOSCI WWW Service!! Date: 10 Dec 1995 23:03:48 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 84 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net I am extremely pleased to announce that BIOSCI is now providing WWW access to all bionet newsgroups through its Web site at www.bio.net!!! To use this service connect to our URL http://www.bio.net and then click on the "Access the BIOSCI/bionet Newsgroups" hyperlink. David Mack, the BIOSCI systems administrator at the U.S. BIOSCI node, has done an excellent job using the hypermail program from Enterprise Integration Technologies to make both current and archived BIOSCI messages available through the World Wide Web. Hypermail converts all e-mail addresses in the messages to mailto: URLs. As long as your Web browser is correctly configured to send e-mail, you can not only read these messages, you can also send either public replies to the newsgroup posting address (present on each message To: line) or private replies to the poster (address on each message's From: line). By selecting the appropriate browser option, you may also be able to include text in your reply from the message to which you are responding. The hypermail archive message headers are threaded by default, but messages can also be displayed chronologically or sorted by author or subject line. This capability gives you, in effect, a threaded newsreader through the Web. All messages posted to the lists are still distributed by e-mail and USENET as usual. However they are now also added to the hypermail archives typically within 20 minutes of posting (a delay was added to avoid file locking problems during periods of heavy use). The new hypermail archive gives you the advantages of USENET without requiring a local news server!! You can keep your personal mail file uncluttered now by using this system instead of e-mail. If you have had problems in the past with new bionet groups not getting established in your local USENET news system, weep no more!! The latest messages will always be here at net.bio.net and all you will need is your Web browser to read and/or post to any of the newsgroups. We will also soon be offering far more WAIS indexes as you will see from browsing the new system. Our goal is to provide, in addition to our current system-wide BIOSCI index, individual indexes for each newsgroup! Although we have done extensive testing, we will not be surprised if users discover some remaining bugs in the system. Please report any problems to us at biosci-help@net.bio.net This new archive also opens up the possibility of sponsorships for individual newsgroups. If your organization is interested in helping support our work through advertising in these WWW forums, i.e., displaying your logo - broadcast ads are not allowed, please contact us at biosci-help@net.bio.net. This is one of the most exciting developments at BIOSCI in quite some time. I want to thank David Mack for all of his hard work getting this system up and running. This entailed many late nights and weekends of effort to configure our system properly. I would also like to thank David States at Washington University for calling this program to our attention several months ago. We had looked at other software for providing Web access to our services and like this one the best. Hypermail was developed by Tom Gruber for Enterprise Integration Technologies (EIT) and later rewritten in C by Kevin Hughes at EIT. It is available for free from EIT. I am also pleased to announce that we have recently developed other software here that makes newsgroup moderation very easy compared to the old methods. We continue to encourage the discussion leaders of the low-to-moderate volume newsgroups to consider moderation to protect your readers against Internet ad spams. Please contact us for details on implementing this. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!chi-news.cic.net!news.math.psu.edu!news.cac.psu.edu!newsserver.jvnc.net!news.caren.net!news.join.ad.jp!news.imnet.ad.jp!ripspost.aist.go.jp!news.tisn.ad.jp!news.nig.ac.jp!labmolg-6!kamuraka From: kamuraka@ddbj.nig.ac.jp (Katsuhiko Murakami) Newsgroups: bionet.molbio.proteins Subject: How to get program "RIBBONS" Date: Sun, 10 Dec 1995 23:11:58 +0900 Organization: National Institute of Genetics Lines: 2 Message-ID: NNTP-Posting-Host: labmolg-6.nig.ac.jp Mime-Version: 1.0 Content-Type: text/plain; charset=iso-2022-jp X-Newsreader: NewsWatcher-J 2.0b19J12 I would like to use biomolecular graphic program "RIBBONS". Please tell me how can I get this program? From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!usenet.eel.ufl.edu!ukma!news.uky.edu!news From: Michael Thompson Newsgroups: bionet.molbio.proteins Subject: Re: ---Please Read This--- Date: 8 Dec 1995 22:28:13 GMT Organization: University of Kentucky Lines: 6 Message-ID: <4aae5t$ph6@service2.uky.edu> References: <4a998n$i6m@luna.vistec.com> NNTP-Posting-Host: 128.163.70.124 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; PPC) X-URL: news:4a998n$i6m@luna.vistec.com Rather than clog up the newsgroups with your ad with the ambiguous title, please identify it as such so that the rest of us who aren't interested can ignore it. Until I read it I thought that it was a serious inquiry for help From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!agate!news.mindlink.net!van-bc!unixg.ubc.ca!hayci.generes.ca!user From: kalch@ulam.generes.ca (Michael kalchman) Newsgroups: bionet.molbio.proteins Subject: Proteins sticking to Glutathione-beads Date: Sun, 10 Dec 1995 22:54:27 -0800 Organization: ubc Lines: 13 Message-ID: NNTP-Posting-Host: hayci.generes.ca Hi proteiners?? We are attempting co-affinity purifications of proteins from tissue culture cells with a GST-fusion protein. However, no matter how much we wash with buffer (including up to 3% NP-40) the proteins like to stick to the beads (or at least to the GST protein) just as much as it likes to stick to the fusion protein. Any help on how to do these damn co-affinity purifications would be FANTASTIC. We are and have been attempting to follow published protocols as well. NOTHING is going correctly....... kalch From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!newsfeed.internetmci.com!in1.uu.net!spcuna!news.columbia.edu!merhaba.cc.columbia.edu!wgf3 From: William Guy Fairbrother Newsgroups: bionet.molbio.proteins Subject: Cell Lines:History Date: Sun, 10 Dec 1995 19:05:23 -0500 Organization: Columbia University Lines: 4 Message-ID: NNTP-Posting-Host: merhaba.cc.columbia.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII X-Sender: wgf3@merhaba.cc.columbia.edu Does anybody know where I could find an historical account of the creation of the HeLa cell line? From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!swrinde!cs.utexas.edu!howland.reston.ans.net!newsserver.jvnc.net!raffles.technet.sg!nova.np.ac.sg!nova.np.ac.sg!news From: 93462036@comet.np.ac.sg (Ang Ching Seng) Newsgroups: bionet.molbio.proteins Subject: Reduction of KLH Date: 8 Dec 1995 11:00:57 +0800 Organization: Ngee Ann Polytechnic, Singapore Lines: 14 Message-ID: <4a89p9$cb0@comet.np.ac.sg> NNTP-Posting-Host: comet.np.ac.sg NNTP-Posting-User: 93462036 Dear people, I'm doing the reduction of KLH to expose the free thiol groups(-SH) for the conjugation of a hapten but i can't seem to reduce any of the KLH. ( I currently using Dithiolthreitol ) Would appreciate very much if anyone could tell me how should i carry it out ( the ratio of protein to DTT i used ranges from 1:50 to 1:15000). THANKS !!!! Ang Ching Seng 93462036@np.ac.sg From owner-proteins@net.bio.net Sat Dec 09 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!newsserver.jvnc.net!raffles.technet.sg!nova.np.ac.sg!nova.np.ac.sg!news From: 93462036@comet.np.ac.sg (Ang Ching Seng) Newsgroups: bionet.molbio.proteins Subject: number of S-S bonds in BSA and KLH Date: 8 Dec 1995 10:47:39 +0800 Organization: Ngee Ann Polytechnic, Singapore Lines: 12 Message-ID: <4a890b$bht@comet.np.ac.sg> NNTP-Posting-Host: comet.np.ac.sg NNTP-Posting-User: 93462036 Dear people, Would appreciate very much if anyone can tell me the therotical number of S-S bonds ( Thiol groups ) in BSA and KLH (Hemocyanin from keyhole limpets ) Thanks!!! Ang Ching Seng 93462036@np.ac.sg From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!swrinde!newsfeed.internetmci.com!btnet!demon!cityscape.co.uk!usenet From: ae55@cityscape.co.uk (John Bates) Newsgroups: bionet.molbio.proteins Subject: Protein-Bound NSAID problem Date: 11 Dec 1995 13:47:23 GMT Organization: IP-GOLD User Lines: 12 Message-ID: <4ahcpb$18b@news.cityscape.co.uk> NNTP-Posting-Host: cisae55.demon.co.uk X-Newsreader: WinVN 0.92.6+ Several of the NSAID drugs are very hydrophobic and readily bind to hydrophobic sites on certain plasma proteins, particularly albumin. This is creating a problem for a flurbiprofen/ibuprofen plasma extraction technique I am using. Any suggestion on how these drugs can be released from these sites. I don't want to use an organic such as methanol. I have used the conventional liquid-liquid and solid phase techniques successfully and in fact they are very simple. The specialised technique I am using however is somewhat different and requires that I "release" the two NSAIDs before extraction. John From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!agate!cocoa.brown.edu!cis-ts1-slip14.cis.brown.edu!user From: Stephen_Lasky@brown.edu (Stephen R. Lasky) Newsgroups: bionet.molbio.proteins Subject: Re: Cell Lines:History Date: Mon, 11 Dec 1995 09:03:48 -0400 Organization: Roger Williams Medical Center/Brown University Lines: 20 Message-ID: References: NNTP-Posting-Host: cis-ts1-slip14.cis.brown.edu X-Newsreader: Value-Added NewsWatcher 2.0b24.0+ In article , William Guy Fairbrother wrote: > Does anybody know where I could find an historical account of the > creation of the HeLa cell line? I'm sorry that I don't know any specific refs to give you but are you any relation to the Dr. Guy (don't recall his first name) who origianally derived the HeLa cell line? SRLasky -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Stephen R. Lasky Ph.D. Brown U/Roger Williams Medical Center, Providence, RI. Phone: 401-456-5672 Fax: 401-456-6569 e:mail: Stephen_Lasky@brown.edu =================================================================== America may be unique in being a country which has leapt from barbarism to decadence without touching civilization. John O'Hara. ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!agate!cgl!espinoza From: espinoza@cgl.ucsf.edu (Hernan Espinoza) Newsgroups: bionet.molbio.proteins Subject: Opinions about Gradifrac? Date: 12 Dec 95 01:03:20 GMT Organization: UCSF Computer Graphics Lab Lines: 4 Message-ID: NNTP-Posting-Host: socrates.ucsf.edu Summary: What do you think of the Gradifrac? Keywords: FPLC Howdy, We are thinking of purchasing a GradiFrac Low Pressure Chromatography System from Pharmacia. Does anyone out there have an opinion/experience with one? My email is espinoza@cgl.ucsf.edu. Thanks in advance! From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!uwm.edu!post.its.mcw.edu!usenet From: "Jennifer L. Potter" Newsgroups: bionet.molbio.proteins Subject: Re: Cell Lines:History Date: 11 Dec 1995 22:22:08 GMT Organization: Medical College of Wisconsin Lines: 23 Message-ID: <4aiaug$di9@post.its.mcw.edu> References: NNTP-Posting-Host: d350-1.biochem.mcw.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) To: wgf3@columbia.edu William Guy Fairbrother wrote: > >Does anybody know where I could find an historical account of the >creation of the HeLa cell line? > Dear William, You may have already checked this source, but I'm posting just in case you have not. The American Type Culture Collection (ATCC) catalog usually gives references documenting the generation of a certain cell line. It had some helpful references for a cell line I was interested in. Other than that, the general knowledge I have about HeLa is that it is derived from a woman named Helen Lane and is a cerivcal cancer. Best of luck, Jennifer Potter Medical College of WI Dept Biochemistry From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!hearst.acc.Virginia.EDU!murdoch!faraday.clas.Virginia.EDU!vk2n From: vk2n@faraday.clas.Virginia.EDU (Vladimir Kalashnikov) Subject: JobNeed-PhD-BioorgChem-Pept/DNA-Syn/Modif X-Nntp-Posting-Host: faraday.clas.virginia.edu Message-ID: Sender: usenet@murdoch.acc.Virginia.EDU Organization: University of Virginia Date: Mon, 11 Dec 1995 17:42:30 GMT Lines: 164 I am writing to inquire about possible positions within R&D departments of chemical, pharmaceutical or biotechnological companies. I am an experienced bioorganic / analytical chemist. CURRICULUM VITAE Vladimir V. KALASHNIKOV, Ph.D. Business address: University of Virginia, Department of Chemistry McCormick Rd. Charlottesville, VA 22901 Phone: (804) 924-3241 Fax: (804) 924-3710 E-mail: vk2n@virginia.edu Home address : 312 13th Street NW Apt. 26 Charlottesville, VA 22903-2754 Phone (804) 984-0571 AREA OF SPECIALIZATION: Bioorganic chemistry. Peptide synthesis (solution, liquid-phase and solid phase), chemistry of side-chain, C- and N-protecting groups, side reactions. Peptide/protein modification and conjugation, synthesis of chromogenic and fluorogenic substrates for enzymes. Chemistry of nucleic acids; oligonucleotide synthesis, modification, crosslinking and conjugation. Crosslinking agents. Bioconjugates. HPLC of biomolecules (analytical and preparative). EDUCATION: 1994- Postdoctoral Fellow with Professor F.A. Etzkorn. University of Virginia, Department of Chemistry, Charlottesville, VA. 1990 Ph.D. in Bioorganic Chemistry Institute of Bioorganic Chemistry, Novosibirsk, Russia 630090 Thesis title: Investigations of base-labile chromogenic C-protecting groups for peptide synthesis in solution. Advisor: Dr. Vladimir V. Samukov 1975-1977 Postgraduate Research Fellow with Professor Z.A. Shabarova Moscow State University, Department of Chemistry, Laboratory of Chemistry of Nucleic Acids, Moscow, Russia 117234 1975 M.S./B.S. in Chemistry (Organic Chemistry) Moscow State University, Department of Chemistry, Moscow, Russia 117234 EMPLOYMENT AND RESEARCH EXPERIENCE (20 years of experimental activity): 1994- Postdoctoral Research Fellow with Professor F.A. Etzkorn, Department of Chemistry, University of Virginia, Charlottesville, VA 22901. Molecular studies of DNA-binding proteins and bent DNA. Oligonucleotide synthesis, DNA modification and crosslinking. Design and synthesis bifunctional crosslinking agents. Oligosaccharide conjugation. HPLC. 1991-1994 Senior Researcher, Head of Laboratory. "Vector-BioProduct" Ltd., Koltsovo, Novosibirsk region, Russia 633159 Preparative peptide synthesis methodology (solution and liquid-phase). Miscellaneous peptides. Major projects: Adrenocorticotropic Hormone, Luotropic Hormone-Releasing Hormone, Oxytocin, Vasopressin and their analogs. Peptides with unnatural amino acids. Substrates for enzymes. HPLC. 1984-1991 Researcher, Senior Researcher, All-Union Research Institute of Molecular Biology. SPU "Vector", Koltsovo, Novosibirsk region, Russia 633159 Peptide synthesis. Liquid-phase synthesis in semi- and preparative scale of neuropeptides, fragments of viral proteins, etc. New protecting groups, new methods of synthesis and deblocking of synthetic peptides, side reactions. Peptide conjugation, "chemical vaccine". HPLC of biomolecules. 1977-1983 Junior Researcher, All-Union Research Institute of Molecular Biology. SPU "Vector", Koltsovo, Novosibirsk region, Russia 633159 Oligonucleotide synthesis (fragments of genes of beta-endorphin, interferons, etc.). Chemistry of nucleosides, synthesis of sorbents for affinity chromatography of DNA and proteins. HPLC of synthetic oligonucleotides and DNA 1975-1977 Postgraduate Research Fellow, Laboratory of Chemistry of Nucleic Acids, Chemistry Department, Moscow State University. Moscow, Russia 117234 Solid- and solution-phase oligonucleotide synthesis; chemistry of nucleic acids AWARDS AND PROFESSIONAL SOCIETIES: 1987 Board of Honor of All-Union Research Institute of Molecular Biology 1978 Ministry of Medical and Microbiological Industry Award 1995 American Chemical Society 1973 Russian Mendeleev Chemical Society REFERENCES A list is available upon request. PUBLICATIONS There are over 25 scientific publications. Publications within the last years (most important): 1. Kalashnikov V.V., Nikolaeva E.N., Polyakova I.M. Synthesis and Deblocking of Peptides, Containing N-(2,4,6-Triisopropylbenzenesulfonyl)Arginine Residuals. Zhurnal Obshchei Khimii, 1994, 64 (4), p. 685-689 2. Kalashnikov V.V., Polyakova I.M. Synthesis of Fragments of Adrenocorticotropic Hormone (ACTH1-10, ACTH11-24 and ACTH1-24) Using Chromogeneous Alkalilabile C-Terminal Protective Group. Zhurnal Obshchei Khimii, 1993, 63 (6), p. 1391-1396 3. Ashmarin I.P., Vakulina O.P., Rozhanetz V.V., Maksimov V.V., Samukov V.V., Kalashnikov V.V., Mizenko G.A. Study of the Seizure Threshold and Increasing Resistance of Rats to Stress After Immunization to Diazepam-Binding Inhibitor Fragment. Bulletin of Experimental Biology and Medicine, 1992, 113 (3), p. 349-352 4. Kalashnikov V.V., Samukov V.V. Alpha-2-Cyanoethyl Ester of tert-Butyloxycarbonyl-Aspartic Acid As an Intermediate Compound for Synthesis of Beta-Esters. Khimiya Prirodnykh Soedinenii, 1990, (2), p. 241-245 5. Kalashnikov V.V., Troshkov M.L., Samukov V.V. Synthesis and Deblocking of Peptides, Containing Beta-Cyclohexylaspartic Acid Residuals. Zhurnal Obshchei Khimii, 1990, 60 (7), p. 1910-1918 6. Shevalier A.F., Samukov V.V., Ofitserov V.I., Kalashnikov V.V., Mizenko G.A., Kolokoltsov A.A. Immunochemical and Biological Properties of Synthetic Fragments from the Amino Acid Sequence-124-144 of Human-Leukocyte Interferon-Alpha-2. Bioorganicheskaya Khimiya, 1990, 16 (7), p. 916-925 7. Mazurkova N.A., Kalashnikov V.V., Mizenko G.A., Samukov V.V., Podchernyaeva R.Y. Study of the Activity of Antibodies to C-Terminal Fragments of Heavy-Chain of Influenza-A (H1-Subtype and H3-Subtype) Virus Hemagglutinins. Voprosy Virusologii, 1990, 35 (4), p. 283-286 8. Petrenko V.A., Kipriyanov S.M., Mizenko G.A., Eroshkin A.M., Sivolobova G.F., Rukavishnikov M.Y., Akimenko Z.A., Boldyrev A.N., Kalashnikov V.V. Construction of a Gene of Hybrid Influenza-Virus Hemagglutinin Subtype-H1-H3 and its Expression in Escherichia Coli. Molecular Biology, 1990, 24 (2), p. 331-339 9. Samukov V.V., Kalashnikov V.V., Shvalie A.F., Ofitserov V.I. Cleavage of 2-Alkylsulfonylethyl and 2-Arylsulfonylethyl Carboxy Protective Groups by Organic Bases - Synthesis of Glutamyl-Containing Fragments of Hemagglutinin of Virus of Flu-A/Leningrad/54/80. Zhurnal Obshchei Khimii, 1989, 59 (3), p. 703-711 10. Kalashnikov V.V., Samukov V.V. Synthesis of C-Terminal Peptide Fragments of a Heavy Chain of Hemagglutinin of H-1 and H-3 Subtype Flu Virus. Khimiya Prirodnykh Soedinenii, 1988, (5), p. 732-738 11. Kalashnikov V.V., Samukov V.V. 2-Cyanoethyl Ester as a New Protecting Group for Carboxyl Function in Peptide Synthesis. Khimiya Prirodnykh Soedinenii, 1988, Iss 3, p. 412-416 12. Samukov V.V., Kalashnikov V.V., Ofitserov V.I., Shvalie A.F. Synthesis of Peptide Fragments of Hemagglutinin of the A/Aichi/2/68 (H3N2) Influenza Virus. Bioorganicheskaya Khimiya, 1985, 11 (8), p. 1037-1047 13. Samukov V.V., Mazurkova N.A., Kalashnikov V.V., Ofitserov V.I., Shvalie A.F., Mizenko G.A., Eroshkin A.M., Podchernyaeva R.Y. Antigenic Properties of Synthetic Fragments of Heavy-Chain of Influenza-A Virus Hemagglutinins. Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 1988, (7), p. 35-39 14. Kalashnikov V.V., Samukov V.V., Shubina T.N., Yamschikov V.F. Application of the Reversed-Phase Chromatography in Oligonucleotide Synthesis. Bioorganicheskaya Khimiya, 1983, 5 (8), p. 666-672 15. Shabarov Yu.S., Mochalov S.S., Fedotov A.N., Kalashnikov V.V. Synthesis of Substituted Anthranils by Deoxidation of o-Nitrosoacylbenzenes. Khimiya Geterotsiklicheskyh Soedinenii, 1975 (9), p. 1195-1197 SCIENTIFIC MEETINGS ATTENDED. (since 1987) 1992 Structure and Function of Regulatory Polypeptides, International Symp., Russia - Germany. Moscow. Presented 1 paper 1990 All-Union Symposium on Chemistry of Peptides, Riga. Presented 1 paper 1987 All-Union Symposium on Chemistry of Proteins and Peptides, Tallinn. Presented 2 papers 1987 All-Union Conference on Methods of Synthesis and Analysis of Biochemical Products, Riga. Presented 1 paper From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!agate!news.ucdavis.edu!library.ucla.edu!newsfeed.internetmci.com!swrinde!cs.utexas.edu!newshost.convex.com!news.duke.edu!galactose.mc.duke.edu.uucp!tschantz From: tschantz@galactose.mc.duke.edu (William P. Tschantz) Newsgroups: bionet.molbio.proteins Subject: Kinase Question Date: 11 Dec 1995 15:14:35 GMT Organization: Duke University; Durham, N.C. Lines: 16 Distribution: world Message-ID: <4ahhsr$ibm@news.duke.edu> NNTP-Posting-Host: galactose.mc.duke.edu Hi I have a question about kinases. Are there any other phosphate donors used by a kinase other than ATP and other nucleotides. I am looking for a enzymatic reation which would use a kinase of some sort to transfer a pyrophosphate group or maybe sequential monophosphate to the substrate. Also maybe examples of phosphate movement other than kinases and the compounds that they use might be helpful. BTW, nucleotides have no effect in my assay. Any help appreciated. Thanks in advance. Bill From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!agate!news.ucdavis.edu!library.ucla.edu!info.ucla.edu!newsfeed.internetmci.com!in1.uu.net!nih-csl!loglady.ninds.nih.gov!johnk From: johnk@spasm.niddk.nih.gov (John Kuszewski) Subject: Re: Protein Models & CAD/CAM Message-ID: <1995Dec11.144209.18678@alw.nih.gov> Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: spasm.niddk.nih.gov Reply-To: johnk@spasm.niddk.nih.gov (John Kuszewski) Organization: National Insts. of Health References: <4a2rre$alp@agate.berkeley.edu> <30C8932B.5862@oci.utoronto.ca> Date: Mon, 11 Dec 1995 14:42:09 GMT Lines: 51 In article <30C8932B.5862@oci.utoronto.ca>, Flip Hoedemaeker writes: |> Louis Hom wrote: |> > |> > It seems to me that life for some folks would be easier if you could have |> > your computer create a real 3D representation of a protein structure. I |> > know that computer-controlled generation of prototypes in manufacturing is |> > done all the time. Does anyone know of someone trying to apply computer |> > aided manufacturing technology to protein structures? |> |> I'm not sure what you mean with "manufacturing technology" , but the SGI's we use |> to model protein structures obtained by X-ray and NMR are exactly the same as what |> car or plane or computer animation designers use. Of course the software is |> different, but we get to see "real" 3D images! |> |> Flip What he means is that engineers now have machines that build real-life, three-dimensional parts out of plastic, straight from a CAD file. These work by either machining down a block of plastic or building one up using UV- sensitive polymerization and a UV laser. And for all that our SGIs can do, there's nothing like holding something in your hand. Unfortunately, there are several drawbacks. The machines are expensive (~$100000) and slow (~1-6 hrs/part). More importantly, they're often not suited to protein structural analysis, since we're often deleting and adding pieces, visualizing interactions, etc. It'd be hard to have a useful protein model that showed both the atoms and a GRASP-style electric field. Proteins are just harder than cars. -- _____________ | ___/_ | |/ / -- /\ // /-- || || / /|| || || / / || || ||/ / || John Kuszewski || |/ /| || johnk@spasm.niddk.nih.gov || / /|| || \/ / / || \/ that's MISTER protein G to you! |/__/| | /_________| "Biophysics has driven me to an attitude of apocalyptic doom" --Frank Delaglio From owner-proteins@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: dinalle@aol.com (Dinalle) Newsgroups: bionet.molbio.proteins Subject: Protein/enzyme purification services available Date: 11 Dec 1995 21:53:47 -0500 Organization: America Online, Inc. (1-800-827-6364) Lines: 25 Sender: root@newsbf02.news.aol.com Message-ID: <4aiqrr$fpv@newsbf02.news.aol.com> Reply-To: dinalle@aol.com (Dinalle) NNTP-Posting-Host: newsbf02.mail.aol.com We have protein or enzyme purification capacity available on a contract basis In house refrigerated centrifugation capabilities include medium (IEC) and large capacity (13L/2500 R.C.F.), as well as 4 Super Sharples (62,800 R.C.F.) continuous flow centrifuges. Coldroom chromatographic capabilities for buffer exchange, size exclusion, ion-exchange and other common separation modes. In addition, staff has long-term experience with PAGE, preparative and analytical isoelectric focusing, FPLC, and other common protein purification and characterization methods. Typical purified end-product amounts are from milligram to gram quantities, depending on source and purity required. Proteins purified have been used for crystallography, basic structure/function studies, etc. Email inquiries or questions to dinalle@aol.com or contact: Alan Bettermann BioRenewal Technologies Inc. Madison, WI Tel 608/ 276-8980 Fax 608/ 273-6989 From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!Germany.EU.net!news.dfn.de!news.coli.uni-sb.de!hades.rz.uni-sb.de!news From: ptpwol@krzsun.med-rz.uni-sb.de (Peter Wollenberg) Newsgroups: bionet.molbio.proteins Subject: Re: Hemoglobin and Cyanide Date: Tue, 12 Dec 1995 14:49:22 GMT Organization: University of Saarland, Computing Center, Germany. Lines: 27 Message-ID: <4ak18r$ksb@hades.rz.uni-sb.de> References: NNTP-Posting-Host: ukh4928.med-pt.uni-sb.de X-Newsreader: Forte Free Agent v0.55 dyanega@denr1.igis.uiuc.edu (Doug Yanega) wrote: >This raises a question: I've been told by a colleague that cyanide builds >up in the body tissues over time, and that after years of slight sublethal >doses, one more tiny dose can "push a person over the edge" suddenly. This >was part of his mandatory education working in an analytical chemistry lab >some 40 years ago. This seems completely at odds with my understanding of >the chemistry involved in cyanide toxicity, and I'm curious as to whether >this is at all possible - I'm a professional entomologist, and am exposed >to sublethal doses of cyanide hundreds of times a year, virtually every >time I use a killing jar. I can't believe that I'm just cranking up some >invisible physiological ratchet, and priming myself to drop dead some day >when I catch that one fatal whiff. Doug, cyanide is converted to the non-toxic and easily excreted thiocyanate by an enzyme called rhodanese in the liver. Therefore, a toxic buildup will not occur. Peter -------------------------------------------------------------- ptpwol@krzsun.med-rz.uni-sb.de P. Wollenberg Dept. of Pharmacology and Toxicology Univ. d. Saarlandes Homburg/Saar Germany From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!nntp.uio.no!ifi.uio.no!news.sics.se!newsfeed.sunet.se!news01.sunet.se!sunic!news99.sunet.se!umdac!news From: Michael Daws Newsgroups: bionet.molbio.proteins Subject: peptide immobilisation Date: 12 Dec 1995 12:10:32 GMT Organization: UCMP Lines: 10 Message-ID: <4ajrfo$m1d@studium.student.umu.se> NNTP-Posting-Host: clsmac.ucmp.umu.se Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; PPC) X-URL: news:bionet.molbio.proteins I have a peptide with a C-terminal cysteine that I want to attach to activated agarose beads. Can anybody suggest a good product for this purpose? My peptide contains a tyrosine, so I don't want to use anything that relies on iodoacetyl chemistry, and also I would prefer the reaction to be irreversible. I then intend to use the immobolised peptide for precipitation from cell lysates, so anything that has high non-specific binding is also out of the question. I would be grateful for any suggestions. From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!EU.net!Norway.EU.net!nntp-oslo.UNINETT.no!nntp-trd.UNINETT.no!nac.no!ifi.uio.no!news.sics.se!newsfeed.sunet.se!news01.sunet.se!sunic!news99.sunet.se!news.funet.fi!mordred.cc.jyu.fi!vap210a.pcbio.jyu.fi!vuento From: vuento@dodo.jyu.fi (Vuento Matti) Newsgroups: bionet.molbio.proteins Subject: Clamydial peptidase Date: Tue, 12 Dec 1995 08:28:52 GMT Organization: Jyv{skyl{n yliopisto Lines: 13 Message-ID: NNTP-Posting-Host: vap210a.pcbio.jyu.fi I have seen claims that Chlamydia trachomatis has a specific peptidase ("peptidase 123") which can be used for clamydial screening. The Medline database seems to know nothing of this. Does anyone have information about clamydial peptidase? Please drop a line in my mailbox. matti vuento vuento@dodo.jyu.fi From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: manjake@aol.com (MANJAKE) Newsgroups: bionet.molbio.proteins Subject: What stimulates interleukin11? Date: 12 Dec 1995 03:58:19 -0500 Organization: America Online, Inc. (1-800-827-6364) Lines: 2 Sender: root@newsbf02.news.aol.com Message-ID: <4ajg7b$otr@newsbf02.news.aol.com> Reply-To: manjake@aol.com (MANJAKE) NNTP-Posting-Host: newsbf02.mail.aol.com Can I stimulate Il11 in the skin? MJ From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!daresbury!not-for-mail From: arturo.galvani@itner.pharmacia.se Newsgroups: bionet.molbio.proteins Subject: Re: Cell Lines:History Date: 12 Dec 1995 08:40:55 -0000 Lines: 14 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <4ajf6n$dl@mserv1.dl.ac.uk> Original-To: proteins@dl.ac.uk I quote from the ATCC catalog: "HeLa was the first aneuploid, epithelial-like cell line to be derived from human tissue and maintained continuosly by serial cell culture. It was isolated by G.O. Gey, W.D. Coffman, and M.T. Kubicek in Febuary, 1951, from a carcinoma of the cervix of a 31 year-old Negro female (Cancer Res. vol. 12, page 254, 1952)." Hope this helps, Arturo Galvani, Pharmacia BioPharm. Nerviano, Milan, Italy. From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!news.euro.net!usenet From: bioup@euronet.nl (Andre W. Schram) Newsgroups: bionet.molbio.proteins Subject: ANNOUNCEMENT: Courses in Protein and Carbohydrate (Bio)technology Date: 12 Dec 1995 19:43:15 GMT Organization: BioUpdate Foundation Lines: 32 Sender: -Not-Authenticated-[8712] Message-ID: <4akm0j$39m@news.euro.net> NNTP-Posting-Host: p011.mas.euronet.nl X-Posted-From: InterNews 1.0.6@p011.mas.euronet.nl Xdisclaimer: No attempt was made to authenticate the sender's name. RESIDENTIAL POST-EXPERIENCE EDUCATION AND TRAINING FOR THE BIOTECHNOLOGY AND ALLIED INDUSTRIES. Established in the Netherlands in 1992, the BioUpdate Foundation promotes residential post-experience education and training for the biotechnology and allied industries. Courses are designed primarily for industrial scientists who are actively involved in protein and carbohydrate (Bio)technology. Courses will also cater for technologists and engineers charged with the development of new products/processes in which proteins and carbohydrates form key elements. Finally, it will help academic and medical researchers and postgraduate students to become familiar with the issues and problems which face those involved in the fast growing "protein and carbohydrate industry". The BioUpdate Foundation also organises workshops and scientific meetings on a selected number of topics related to the areas of protein and carbohydrate (bio)technology. First course to come: "Modules in Biotechnology", to be held March 1996 in the Netherlands. ------------------------------------------------------------------------ - For details on events, please visit the BioUpdate Homepage at: http://www.euronet.nl/users/bioup or e-mail to: bioup@euronet.nl From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!EU.net!peer-news.britain.eu.net!lyra.csx.cam.ac.uk!news From: Peter Wang Newsgroups: bionet.molbio.proteins Subject: Re: origin of HeLa cells Date: 12 Dec 1995 20:04:19 GMT Organization: Centre for Protein Engineering Lines: 26 Message-ID: <4akn83$3no@lyra.csx.cam.ac.uk> References: <4aiaug$di9@post.its.mcw.edu> NNTP-Posting-Host: macx45.mrc-lmb.cam.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) X-URL: news:4aiaug$di9@post.its.mcw.edu >William Guy Fairbrother wrote: >> >>Does anybody know where I could find an historical account of the >>creation of the HeLa cell line? "Jennifer L. Potter" wrote: >...Other than that, the general knowledge I have about HeLa is that >it is derived from a woman named Helen Lane and is a cerivcal cancer. I thought her name was Henrietta Lacks, ring a bell with anyone else? I read an article many years ago about the history of the HeLa cell line (and how it notoriously contaminated other cell cultures so that people ended up working with HeLa rather than the tissue type they thought they were working on). I can't remember where though (maybe Scientific American?) so that's no help. --------------------------------------------------------- Peter Wang, M.D., Ph.D. MRC Centre for Protein Engineering, Hills Road, Cambridge, CB2 2QH, England Tel (01223) 402100 (international calls +44-1223-402100) (01223) 402104 (direct number) Fax (01223) 402140 --------------------------------------------------------- From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!chi-news.cic.net!nntp.coast.net!zombie.ncsc.mil!cs.umd.edu!haven.umd.edu!purdue!lerc.nasa.gov!magnus.acs.ohio-state.edu!infoserver.bgsu.edu!usenet From: Charles Ransam Babu Newsgroups: bionet.molbio.proteins Subject: Re: How to get program "RIBBONS" Date: 13 Dec 1995 01:09:40 GMT Organization: Center for Photochemical Sciences Lines: 7 Message-ID: <4al94k$375@infoserver.bgsu.edu> References: NNTP-Posting-Host: m254-152.bgsu.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) To: kamuraka@ddbj.nig.ac.jp X-URL: news:kamuraka-1012952311580001@labmolg-6.lab.nig.ac.jp Hi! Check out: http://www.cmc.uab.edu:8001/www/ribbons/ribbons.html. I think, you need to buy the license. Best regards, Charles From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!EU.net!sun4nl!news.nic.surfnet.nl!news.wau.nl!usenet From: Javier Ramos Newsgroups: bionet.molbio.proteins Subject: Problems with Streptavidine-HRP Date: 12 Dec 1995 17:11:53 GMT Organization: Wageningen Agricultural University Lines: 21 Message-ID: <4akd4p$p9v@Trex.IenD.wau.nl> NNTP-Posting-Host: dvandusschoten2.mf.wau.nl Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) X-URL: news:bionet.molbio.proteins After getting "too many bands" in a western-blot where I used monoclonal anti-HA antibody coupled to biotin, I made a rutinary essay with only Streptavidine-Horse radish peroxidase and no antibody (I use Boehringer chemiluminiscence kit). Even after lowering the concentration of Strp-HRP to 1 to 20000 I got almost the same pattern of bands. Then trying to avoid biotin, I tested anti-rabbit goat antibody coupled to HRP (I have also a rabbit policlonal anti-HA), this time just in dot blots (no first antibody) and still I get clear "positives" at low concentration. My samples are plants (Vicia hirsuta) extracts, are these common problems? Do you know the explanation? Thank you. Javier Ramos Department of Molecular Biology Wageningen Agricultural University The Netherlands From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!EU.net!peer-news.britain.eu.net!warwick!bham!bhamcs!news.ox.ac.uk!biomac7.jr2.ox.ac.uk!vurquidiI From: vurquidiI@molbiol.ox.ac.uk (Virginia Urquidi) Newsgroups: bionet.molbio.proteins Subject: Re: Gel-elution Date: Tue, 12 Dec 95 20:00:38 GMT Organization: Oxford University Lines: 26 Message-ID: References: <1995Nov27.134900@crick.rz-berlin.mpg.de> NNTP-Posting-Host: biomac7.jr2.ox.ac.uk X-Newsreader: VersaTerm Link v1.1.6 In Article <1995Nov27.134900@crick.rz-berlin.mpg.de>, fester@mpimg-berlin-dahlem.mpg.de wrote: >I would like to elute protein complexes (mitochondrial respiratory >complex III) from Blue Native Gels (about 7% acrylamide) I was considering to recover a protein from a gel using either 1) the BAC crosslinker (Bio-Rad Cat 161-0204) and dissolve the gel with B-merc.etoh or DTT (the protein must contain no disulfide bonds) or 2) from a slab Sea Plaque agarose gel melted and digested with B-Agarase. (FMC Bioproducts) The remaining agarose oligosaccharides could be removed by filtration through membranes of defined MW cut-off (Millipore, Amicon, Flowgen) But I think that the reverse electrophoresis method posted by H Roychowdhury is a very good idea. Best luck Virginia Urquidi vurquidi@molbiol.ox.ac.uk From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!news.ecn.bgu.edu!newspump.wustl.edu!darwin.sura.net!blaze.cs.jhu.edu!RacerX.mse.jhu.edu!news.jhu.edu!NewsWatcher!user From: marco_muzi-falconi@qmail.bs.jhu.edu (marco muzi falconi) Newsgroups: bionet.molbio.proteins Subject: GST antibodies Date: 12 Dec 1995 22:05:18 GMT Organization: jhu Lines: 7 Message-ID: NNTP-Posting-Host: 128.220.123.38 X-Newsreader: Value-Added NewsWatcher 2.0b22.0+ Has anybody out there tried a commercial source of GST antibodies? I need them for Western blotting. I know that SIGMA sells crude serum and also monoclonals. PHARMACIA sells polyclonals as part of a kit. I would like to find out if anybody had good (or bad) experience with them. thanks From owner-proteins@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!noc.near.net!das-news2.harvard.edu!fas-news.harvard.edu!fas.harvard.edu!rickles From: rickles@fas.harvard.edu (Richard Rickles)