From owner-7tms_r@net.bio.net Sun Jun 02 23:00:00 1996 Path: biosci!rutgers!uwm.edu!chi-news.cic.net!newsfeed.internetmci.com!newsserver.jvnc.net!netnews.sbphrd.com!Steven_McClue-1 From: Steven_McClue-1@sbphrd.com (Steve McClue) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Will 7tms survive SDS-PAGE? Date: Mon, 03 Jun 1996 11:26:28 +0000 Organization: Smithkline Beecham Lines: 13 Message-ID: NNTP-Posting-Host: ham992.ha.uk.sbphrd.com X-Newsreader: Yet Another NewsWatcher 2.2.0b6 Does anyone know if it's possible to separate a (peptide) 7TM from other proteins using SDS-PAGE and still retain radioligand binding activity? I'm wondering if I can get at my receptor by SDS-PAGE, western-blotting and then ligand binding. If you know that this is feasible, could you post a reference? Thanks, Steve -- My opinions, not my employer's. From owner-7tms_r@net.bio.net Sun Jun 02 23:00:00 1996 Path: biosci!rutgers!uwm.edu!chi-news.cic.net!newsxfer2.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Will 7tms survive SDS-PAGE? Date: Mon, 03 Jun 1996 16:37:03 -0400 Organization: University of Michigan Lines: 24 Message-ID: <31B34CEF.68F2@umich.edu> References: Reply-To: RNeubig@umich.edu NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0b4Gold (Win95; I) To: Steve McClue Steve, I'd say that you are unlikely to be successful with this but I've learned never to say "It will never work". Why not use a photoaffinity label? Rick RNeubig@umich.edu http://www-personal.umich.edu/~rneubig Steve McClue wrote: > > Does anyone know if it's possible to separate a (peptide) 7TM from other > proteins using SDS-PAGE > and still retain radioligand binding activity? I'm wondering if I can get > at my receptor by SDS-PAGE, > western-blotting and then ligand binding. If you know that this is > feasible, could you post a reference? > > Thanks, > > Steve > > -- > My opinions, not my employer's. From owner-7tms_r@net.bio.net Mon Jun 03 23:00:00 1996 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!rutgers!uwm.edu!newsfeed.internetmci.com!in1.uu.net!hearst.acc.Virginia.EDU!murdoch!usenet From: Bruce Gaylinn Subject: Re: Will 7tms survive SDS-PAGE? X-Nntp-Posting-Host: bootp-44-167.bootp.virginia.edu Content-Type: text/plain; charset=us-ascii Message-ID: <31B427F3.3300@Virginia.edu> Sender: usenet@murdoch.acc.Virginia.EDU Content-Transfer-Encoding: 7bit Organization: Endocrinology References: Mime-Version: 1.0 Date: Tue, 4 Jun 1996 12:11:31 GMT X-Mailer: Mozilla 3.0b4 (Win95; I) Lines: 6 The general answer is no, I am not aware of any "normal" GPCR surviving this. The exception, as I understand it, is the glycoprotein receptors, (LH, FSH, TSH) where the N-terminal extracellular domain is essentially a high affinity binding protein and does function in ligand blots. Bruce From owner-7tms_r@net.bio.net Tue Jun 04 23:00:00 1996 Path: biosci!BIOCELL.IRMKANT.RM.CNR.IT!7tm From: 7tm@BIOCELL.IRMKANT.RM.CNR.IT (Transmembrane receptors) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Review GPCR-Antibody Production Date: 5 Jun 1996 02:48:06 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hello Lena here is the URL address of a very comprehensive list of "antibody-related" WEB sites: http://www-chem.ucsd.edu/Faculty/goodman/antibody.html/abpage.html Regards Raffaele Matteoni ------- From owner-7tms_r@net.bio.net Thu Jun 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!newsfeed.internetmci.com!globe.indirect.com!s71.phxslip4.indirect.com!user From: mthompson@asu.edu (M Thompson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Attachment method for chromophore onto a protein Date: 8 Jun 1996 00:15:19 GMT Organization: Arizona State University Lines: 8 Message-ID: NNTP-Posting-Host: s71.phxslip4.indirect.com I am looking for methods to attach a chromophore, such as thiazole orange and yellow, to a cysteine at residue 1 via a disulfide bridge. One idea is the disufyl-dipyridyl activation of either the chromophore or the protein, with subsequent covalent bond formation of the other. I was wondering if there is an individual who may have experimented with this reaction or has thoughts on this approach or possibly another method. Thanks in advance. From owner-7tms_r@net.bio.net Tue Jun 11 23:00:00 1996 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: IMPORTANT - BIOSCI Fundraising Update! Date: 12 Jun 1996 02:00:32 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 154 Sender: daemon@net.bio.net Distribution: world Message-ID: <199606120900.CAA02967@net.bio.net> NNTP-Posting-Host: net.bio.net BIOSCI is about halfway to its funding goal!! I'm interrupting the usual monthly posting of the BIOSCI miniFAQ to bring you up to date on BIOSCI fundraising progress, a topic of concern to your future use of this resource. Thank you in advance for taking the time to read this message carefully. Last year we announced that BIOSCI was going to adopt the U.S. Public Broadcasting System model to fund its operations after our DOE/NSF grant runs out later this year. Unlike PBS, we are not soliciting contributions from users; we are only selling ads on our Web pages solely to cover our operating costs. Our goal is to seek sponsorships until we build up an operating reserve of about $100,000 and then cease further promotions until we need to build the reserve back up. (The accountants among our readership will be familiar with the problem of deferred revenue which we can not safely utilize until ads have been displayed for a period of time.) We are only about halfway to our funding goal and need to raise further funds to avoid having to curtail services at net.bio.net. Fundraising is time-consuming, however, and we need your help as explained further below. Our operating costs consist of our network connection, phone lines, hardware maintenance (we will be getting newer and faster hardware soon!), plus 0.7 FTE of salaries covering UNIX systems admin, technical support, quality assurance, i.e., testing, of our system, and administrative costs (such as the time it takes to actually find/write/call potential sponsors and raise money!). Although the BIOSCI staff does get compensated for a portion of the work that they do, this project has always received a lot of free after-hours and "vacation" time labor, so we hope that no one will begrudge the time that we do charge to the project to serve you. All of the three part-time staff members, Dave Mack, Julie Lawrence, and myself, have full time day jobs and families in addition to working hard to keep this service running for all of you. Julie and Dave Mack are subcontractors for BIOSCI; my time that is charged to the project defrays a portion of my regular salary instead of adding to my income. Besides having to relocate the project, we were very busy this last year building new infrastructure such as our WWW hypermail interface to the system. This was released last December along with scores of WAIS indices for the newsgroups. Virtually everything is complete, although we do continue to find and fix bugs (many through your helpful feedback!). We are still having some problems with our WAIS indexing. The archives continue to grow rapidly. We are running over 100 indexes now versus three previously and any systems crashes cause greater havoc with the indexing than before! We are still working to fix this as fast as our resources permit and appreciate your patience, but we have been able to automate a lot of the infrastructure to reduce labor as compared to past requirements. We have also implemented new software to make moderation of BIOSCI/bionet newsgroups much easier and combat the growing problem of Internet junk mail and USENET "spamming." About 20% of our groups are now moderated, many of them by the BIOSCI staff! This, for example, made a major difference last year in the quality of content in our EMPLOYMENT/bionet.jobs.offered newsgroup which many commercial concerns and recruiting firms are using **without charge** to recruit candidates for positions in the biological sciences. We are also now in a position to have sponsors for individual newsgroups as you will have noticed if you have visited http://www.bio.net/ and clicked on "Access the BIOSCI/bionet newsgroups" recently. So, how can you help?? ---------------------- As noted above it can take a lot of time to contact potential sponsors if I have to do it all myself. Our request is quite simple. You can do two important things which will take very little time for you individually. First, please use our WWW system at http://www.bio.net/ to access the archives. You can now post or reply to messages via your Web browser. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. Our hope is to quickly raise several large corporate/institutional sponsors on our heavily-used WWW locations (some stats appended below), and then end this sponsorship campaign so that our resources can continue to be used for service provision, not fundraising. Many of our specialty newsgroup WWW archives are still used by small communities of scientists (and they haven't been heavily promoted yet). While these may be valuable niche markets to some advertisers, it will generate more labor and overhead having to find these sponsors, fairly price the locations, and deal with lots of smaller sponsorships than fewer mid-to large sponsors. We are striving to keep our operation as lean and efficient as possible since we are not trying to make careers out of running BIOSCI. We are trying if at all possible to avoid the administrative overhead entailed with processing lots of small payments to reach our fundraising goals. I'd like to thank all of you for your help in advance. In helping us, you are also helping yourselves, not only in keeping this resource available for all of the both large and small research communities that we serve, but also by alleviating the need for us to go back and compete with researchers for tight grant dollars! We promised NSF when we were awarded the BIOSCI grant that we would carry out this mission to make the service self-supporting. With your help, we will succeed in continuing BIOSCI's work into its second decade. Thank you very much! Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net A list of our prime WWW sponsorship locations follow. Please contact us for further details. ---------------------------------------------------------------------- The overall BIOSCI WWW pages are currently visited by users from close to 5500 unique computer hosts per week. Web servers only log the Internet computer/host name and frequently more than one individual can connect to us from a particular host. Main home page, http://www.bio.net, visited recently by about 2100 unique hosts per week Main Newsgroups archives page, http://www.bio.net/archives.html, visited recently by about 1200 Unique hosts per week BIO-JOURNALS archive page, http://www.bio.net/BIO-JOURNALS.html, visited recently by about 1000 unique hosts per week. EMPLOYMENT archive pages: http://www.bio.net:80/hypermail/EMPLOYMENT/ and monthly header pages, visited recently by about 800 unique hosts per week. Address database search page, http://www.bio.net/addrsearch.html, visited recently by about 450 unique hosts per week. Methods newsgroup archive pages, http://www.bio.net:80/hypermail/METHDS- REAGNTS/ and monthly header pages, visited recently by about 350 unique hosts per week. Ads can also be displayed on various combinations of other BIOSCI/bionet newsgroups. Please contact us at biosci-help@net.bio.net for details. ---------------------------------------------------------------------- From owner-7tms_r@net.bio.net Wed Jun 12 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!globe.indirect.com!s82.phxslip4.indirect.com!user From: mthompson@asu.edu (M Thompson) Newsgroups: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.proteins.fluorescent Subject: Synthesis method for thiazole orange Date: 13 Jun 1996 20:38:19 GMT Organization: Arizona State University Lines: 6 Message-ID: NNTP-Posting-Host: s82.phxslip4.indirect.com Xref: biosci bionet.molbio.proteins:8127 bionet.molbio.proteins.7tms_r:741 bionet.molbio.proteins.fluorescent:532 Does anyone have or know the synthesis method for thiazole orange? Any pointers? I have a method for the synthesis, but it leaves out valuable information. If anyone can help me narrow down the amounts, quantities and general missing bits of the reactions involved I will be happy to hear from you. Thanks in advance. From owner-7tms_r@net.bio.net Thu Jun 13 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!vixen.cso.uiuc.edu!tgevax.life.uiuc.edu!saurav From: saurav@tgevax.life.uiuc.edu (Saurav Misra) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Attachment method for chromophore onto a protein Date: 14 Jun 96 03:35:12 GMT Organization: University of Illinois at Urbana Lines: 43 Message-ID: References: NNTP-Posting-Host: tgevax.life.uiuc.edu mthompson@asu.edu (M Thompson) writes: >I am looking for methods to attach a chromophore, such as thiazole orange >and yellow, to a cysteine at residue 1 via a disulfide bridge. One idea is >the disufyl-dipyridyl activation of either the chromophore or the protein, >with subsequent covalent bond formation of the other. I was wondering if >there is an individual who may have experimented with this reaction or has >thoughts on this approach or possibly another method. >Thanks in advance. Although I am admittedly a novice, aren't there already chromophore derivatives being made with reactive thioester groups? I know Molecular Probes sells a bunch of thioester flourescein derivaties. Here we go, I'm quoting directly from the Molecular Probes handbook: "In most cases, a thiol reacts with an alkylating group (R'-X) to yield a very stable tbioether (R-S-R'). The leaving group X may be Cl, Br, I , a sulfonate ester or part of an aziridine ring. Certain other reagents such as NBD chloride react with thiols or amines by a similar substitution of the aromatic chloride. In the case of maleimides and derivatives of acrylic acid, the reaction involves addition of the thiol (R-SH) across the double bond (-C=C-) of th eprobe to again give a thioether (R-S-CHCH-). The only other thiol-reactive reagents offered are those that undergo a thiol-disulfied (R'-S-S-R') interchange reaction to give a new assymetric disulfide (R-S-S-R'). This reaction is freely reversible and thiol-specific." OK, so I didn't know what I was talking about with the thioester junk up top. But looking further into the Molecular probes handbook, I see that they sell a bunch of fluoresent probes, including various iodacetamide derivatives. Many researchers who work on bacteriorhodopsin have used cystine-labeling methods to attach ph-sensitive dyes : see papers from the past few years of Alexiev, Heyn, and Khorana for some experimental techniques on how to do this. It looks fairly straightforward. If you don't have access to the Molecular Probes Handbook, e-mail me I I will fax you the relevant pages. The company itself can be reached at (503) 465-8353 (Tech. assistance) or -8338 (Customer service). They should be willing to give advice. -Saurav Misra Univ of Illinois / Biophysics saurav@tgevax.life.uiuc.edu From owner-7tms_r@net.bio.net Tue Jun 18 23:00:00 1996 Path: biosci!CHEM.VU.NL!vheerik From: vheerik@CHEM.VU.NL (Harm van Heerikhuizen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: SD on Ki or on pKi? Date: 19 Jun 1996 01:23:29 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 26 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear Netters, We are a group of molecular biologists working on GPCRs, and accidently we ended up performing pharmacological experiments. Having carried out an extensive pharmacological characterization of a number of GPCRs cloned by us we encountered the following problem. What is the proper way to present the data of, for instance, the displacement of a radioligand by (ant)agonists? (i) as a Ki-value +/- SD or (ii) as a pKi-value +/- SD We have noticed that both representations are being used in the literature. Obviously only one can be correct. Which one, and why?? Thanks Harm van Heerikhuizen Molecular Neurobiology group Dept. of Biochemistry & Mol. Biol. Vrije Universiteit, Amsterdam The Netherlands From owner-7tms_r@net.bio.net Tue Jun 18 23:00:00 1996 Path: biosci!UNMCVM.UNMC.EDU!DBYLUND From: DBYLUND@UNMCVM.UNMC.EDU ("David B. Bylund --UNMCVM", DBYLUND) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ki or pKi Date: 19 Jun 1996 09:42:27 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 33 Sender: daemon@net.bio.net Distribution: world Message-ID: <199606191642.JAA22150@net.bio.net> NNTP-Posting-Host: net.bio.net FROM: David B. Bylund --UNMCVM(DBYLUND) Harm van Heerikhuizen wrote asking about the proper way to present the data for the inhibition (not displacement) of radioligand binding by (ant)agonists? (I) as a Ki-value +/- SD or (ii) as a pKi-value +/- SD The data can be correctly presented either way. Personally, I think in Ki values, not pKi values, so I prefer Ki. What is important , however, is how the data are calculated. It is generally accepted that Ki values are log normally distributed, and thus the mean and SE should be calculated from the pKi values, The mean and SE can then be converted back to Ki values. Thus, the Ki values are reported as geometric means +/-S.E. Converting from the mean pKi to the mean Ki value is no problem. For the SE the procedure that is generally used is to multiply the mean Ki value by the SE of the pKi mean. See DeLean, A., Hancock, A.A., & Lefkowitz, R.J. (1982). Validation and statistical analysis of a computer modeling method for quantitative analysis of radioligand binding data for mixtures of pharmacological receptor subtypes. Mol Pharmacol, 21, 5-16. David B. Bylund, Ph.D. Department of Pharmacology University of Nebraska Medical Center P.O. Box 986260 Omaha, NE 68198-6260 voice: 402/559-4788 fax: 402/559-7495 EMAIL: DBYLUND@UNMC.EDU From owner-7tms_r@net.bio.net Wed Jun 19 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!news.uit.no!atf1!edvard From: edvard@atf1.imb.fm.uit.no (Oyvind Edvardsen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Mutant database GRAP unavailable Date: 20 Jun 1996 09:04:08 GMT Organization: University of Tromsoe Lines: 26 Distribution: world Message-ID: <4qb468$m6e@news.uit.no> Reply-To: edvard@fagmed.uit.no NNTP-Posting-Host: atf1.imb.fm.uit.no Due to a crash on our server system disk, GRAP has been unavailable for some days (you have probably noticed by now). We are currently slowly recovering, but GRAP will still be unavailable for a few more days. When being struck by hardware problems, we take the opportunity to do some hardware update/reorganization and software/OS updates. Thus, we will still need some more time to be back in business. We expect GRAP to become available during the upcoming weekend or shortly after. Details will be posted later. Sincerely, Dr. OEyvind Edvardsen -o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o- ___ _ | /| _| |_ _| __ _ _| _ _ |/_| \/ \/ | |\| |_| |_ |_| \/ |_|_ | |_| _\ |_' |\| / _____________________________________________________________________________ School of Medicine | Dept. of Pharmacology, IMB | TelePhone: +47 77 64 53 42 University of Tromsoe | TeleFax: +47 77 64 53 10 MH, Breivika | Email: edvard@fagmed.uit.no N-9037 TROMSOE, NORWAY | URL: http://atf1.fagmed.uit.no/mgl.html ------------------------------------------------------------------------------ From owner-7tms_r@net.bio.net Thu Jun 20 23:00:00 1996 Path: biosci!rutgers!uwm.edu!chi-news.cic.net!news.compuserve.com!news.production.compuserve.com!news From: DSF <76042.532@CompuServe.COM> Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.yeast Subject: antibodies to yeast proteins Date: 21 Jun 1996 19:48:04 GMT Organization: Cadus Lines: 3 Message-ID: <4qeu9k$a7$1@mhade.production.compuserve.com> Xref: biosci bionet.molbio.methds-reagnts:46078 bionet.molbio.proteins:8177 bionet.molbio.proteins.7tms_r:746 bionet.molbio.yeast:5439 I'm looking for commercially available antibodies (preferably monoclonals but I'm not too picky) to ubiquitously expressed yeast proteins. Suggestions? From owner-7tms_r@net.bio.net Fri Jun 21 23:00:00 1996 Path: biosci!OYSTER.SMCM.EDU!rmyerowi From: rmyerowi@OYSTER.SMCM.EDU (Rachel Myerowitz) Newsgroups: bionet.molbio.proteins.7tms_r Subject: text for molecular genetics of eukaryotes Date: 22 Jun 1996 21:26:37 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Does anybody have any suggestions for a text for a seminar course concerned with the molecular genetics of eukaryotes? The course meets once a week for 2 hours for 14 weeks. It is an undergraduate class. There will be twelve students in the class and the students will have mixed backgrounds-some have had a lab class concerned with recombinant DNA methods and others have lttle background.There will be no lab component. I would like to discuss organizatiion, complexity and flexibility of the genome as well as regulation of gene expression. I would also like to discuss in vitro and in vivvo systems for studying gene expressions like transgeneics and targeted gene disruption. Any suggestions would be greatly appreciated. Rachel Myerowitz- email address is rmyerowi@oyster.smcm.edu From owner-7tms_r@net.bio.net Sat Jun 22 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!quagga.ru.ac.za!uct.ac.za!news From: david@chempath.uct.ac.za (Dave Myburgh) Newsgroups: bionet.molbio.proteins.7tms_r Subject: GTP gamma S binding assay method? Date: Sun, 23 Jun 1996 14:23:18 GMT Organization: UCT Medical School Lines: 11 Message-ID: <4qjjnu$95a@groa.uct.ac.za> NNTP-Posting-Host: med52.med.uct.ac.za X-Newsreader: Forte Agent .99b.112 Hi All Does anyone out there have a method for doing GTP gamma S binding assays on membranes of GPCRs expressed in COS-1 cells (or any similar cell line). Thanks in advance Dave From owner-7tms_r@net.bio.net Sun Jun 23 23:00:00 1996 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!galaxy.ucr.edu!ihnp4.ucsd.edu!munnari.OZ.AU!news.mel.connect.com.au!news.mira.net.au!harbinger.cc.monash.edu.au!nntp.coast.net!howland.reston.ans.net!gatech!usenet.eel.ufl.edu!warwick!bsmail!usenet From: "Dr Andrew J. Doherty" Subject: Re: GTP gamma S binding assay method? Content-Type: multipart/mixed; boundary="-------------------------------257192377724860" Message-ID: To: david@chempath.uct.ac.za Sender: usenet@uns.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: anaajd.ana.bris.ac.uk Organization: University of Bristol, UK References: <4qjjnu$95a@groa.uct.ac.za> Mime-Version: 1.0 Date: Mon, 24 Jun 1996 08:06:42 GMT X-Mailer: Mozilla 1.1N (Windows; I; 16bit) Lines: 16 This is a multi-part message in MIME format. ---------------------------------257192377724860 Content-Transfer-Encoding: 7bit Content-Type: text/plain; charset=us-ascii I'd also be very interested in such a methodology for CHO cells Ta very much Andy Doherty ---------------------------------257192377724860 Content-Transfer-Encoding: 7bit Content-Type: ---------------------------------257192377724860-- From owner-7tms_r@net.bio.net Sun Jun 23 23:00:00 1996 Path: biosci!galaxy.ucr.edu!ihnp4.ucsd.edu!swrinde!cs.utexas.edu!howland.reston.ans.net!newsfeed.internetmci.com!news.ac.net!news.cais.net!nntp.uio.no!nntp-oslo.UNINETT.no!nntp-trd.UNINETT.no!news.uit.no!atf1!edvard From: edvard@atf1.imb.fm.uit.no (Oyvind Edvardsen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Mutant database GRAP up and running Date: 24 Jun 1996 09:43:48 GMT Organization: University of Tromsoe Lines: 22 Distribution: world Message-ID: <4qlo0k$pdb@news.uit.no> Reply-To: edvard@fagmed.uit.no NNTP-Posting-Host: atf1.imb.fm.uit.no GRAP is now in normal operations again. There may, however, still be some small instabilities. Thanks for your patience! Sincerely, Dr. OEyvind Edvardsen -o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o- ___ _ | /| _| |_ _| __ _ _| _ _ |/_| \/ \/ | |\| |_| |_ |_| \/ |_|_ | |_| _\ |_' |\| / _____________________________________________________________________________ School of Medicine | Dept. of Pharmacology, IMB | TelePhone: +47 77 64 53 42 University of Tromsoe | TeleFax: +47 77 64 53 10 MH, Breivika | Email: edvard@fagmed.uit.no N-9037 TROMSOE, NORWAY | URL: http://atf1.fagmed.uit.no/mgl.html ------------------------------------------------------------------------------ From owner-7tms_r@net.bio.net Mon Jun 24 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!newsserver.jvnc.net!netnews.sbphrd.com!ham180.ha.uk.sbphrd.com!user From: Tim_Young-1@sbphrd.com (Tim Young) Newsgroups: bionet.molbio.proteins.7tms_r Subject: [33P]-alpha-ATP Date: 25 Jun 1996 11:31:24 GMT Organization: SmithKline Beecham Lines: 7 Message-ID: Reply-To: Tim_Young@sbphrd.com NNTP-Posting-Host: ham180.ha.uk.sbphrd.com X-Newsreader: Value-Added NewsWatcher 2.0b22.0+ Hi, I'm looking for a source of [33P]-alpha-ATP for a cyclase assay, can anybody help. Cheers, ;-) -- The opinions expressed in this communication are my own, and do not necessarily reflect those of my employer. From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!chi-news.cic.net!newsfeeder.sdsu.edu!newspump.sol.net!homer.alpha.net!news.ultranet.com!usenet From: "John P. McGrath" Newsgroups: bionet.molbio.proteins.7tms_r Subject: BK receptor antibody search Date: Wed, 26 Jun 1996 15:06:37 -0400 Organization: UltraNet Communications, Inc. Lines: 13 Message-ID: <31D18A3D.41@ma.ultranet.com> Reply-To: Cambridge, MA NNTP-Posting-Host: 146.115.77.18 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.02 (Macintosh; I; PPC) I'm in search of antibodies to the rat bradykinin B2 receptor for use in immunohistochemical studies. Any information re: availability of such antibodies and their use would be greatly appreciated. Thanks in advance, John McGrath Alkermes, Inc. 64 Sidney Street Cambridge, MA 02139 bigred@ma.ultranet.com From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!online.no!Norway.EU.net!EU.net!news-res.gsl.net!news.gsl.net!usenet.eel.ufl.edu!usenet.cis.ufl.edu!caen!newsxfer2.itd.umich.edu!newsfeed.internetmci.com!uwm.edu!news.cse.psu.edu!news.eecs.nwu.edu!newsfeed.acns.nwu.edu!news.acns.nwu.edu!NewsWatcher!user From: VENITA@nwu.edu (VENITA DE ALMEIDA) Newsgroups: bionet.molbio.proteins.7tms_r Subject: program for analysis of ligand binding FOR MACINTOSH Date: Tue, 25 Jun 1996 18:33:55 -0500 Organization: nu Lines: 8 Message-ID: NNTP-Posting-Host: 129.105.41.196 does anyone know where i can get the program for analysis of ligand binding? There is aversion of MAC Ligand on the web but it doesn't work on my computer. Would appreciate any information. Thanks venita I DID NOT SPECIFY IN MY EARLIER MESSAGE THAT IT WAS FOR USE ON MACINTOSH OR POWER PC From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!news.compuserve.com!news.production.compuserve.com!news From: DSF <76042.532@CompuServe.COM> Newsgroups: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.yeast Subject: Yeast antibodies!! Date: 25 Jun 1996 21:18:32 GMT Organization: Cadus Lines: 4 Message-ID: <4qpl38$kt3$1@mhadf.production.compuserve.com> Xref: biosci bionet.molbio.proteins:8190 bionet.molbio.proteins.7tms_r:754 bionet.molbio.yeast:5453 I'm desperately looking for commercially available antibodies to ubiquitously expressed yeast proteins, for instance, anti-TATA Binding Protein. HELP!! Lynette From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!cdc2.cdc.net!vixen.cso.uiuc.edu!uchinews!news From: Kelly Ambler Subject: Re: Program for analysis of ligand binding data 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: Mime-Version: 1.0 Date: Tue, 25 Jun 1996 21:05:14 GMT X-Mailer: Mozilla 1.1N (Windows; I; 16bit) Lines: 6 Several programs are available commercially. Our lab uses the GraphPad Prism program. (no affiliation, etc.) Kelly Ambler University of Chicago From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Path: biosci!daresbury!bioftp.unibas.ch!infobiogen.fr!jussieu.fr!oleane!tank.news.pipex.net!pipex!howland.reston.ans.net!gatech!usenet.eel.ufl.edu!news.ultranet.com!homer.alpha.net!uwm.edu!news.cse.psu.edu!news.eecs.nwu.edu!newsfeed.acns.nwu.edu!news.acns.nwu.edu!NewsWatcher!user From: venita@nwu.edu (Venita De Almeida) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Program for analysis of ligand binding data Date: Tue, 25 Jun 1996 11:32:12 -0500 Organization: nu Lines: 5 Message-ID: NNTP-Posting-Host: 129.105.41.196 does anyone know where i can get the program for analysis of ligand binding? There is aversion of MAC Ligand on the web but it doesn't work on my computer. Would appreciate any information. Thanks venita From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!news.ycc.yale.edu!news From: Carsten Schubert Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: GTP gamma S binding assay method? Date: Wed, 26 Jun 1996 16:41:52 -0400 Organization: Yale University Lines: 28 Message-ID: <31D1A090.145B@csb.yale.edu> References: <4qjjnu$95a@groa.uct.ac.za> NNTP-Posting-Host: sabbath.csb.yale.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.01 (Macintosh; I; PPC) Dave Myburgh wrote: > > Hi All > > Does anyone out there have a method for doing GTP gamma S binding > assays on membranes of GPCRs expressed in COS-1 cells (or any similar > cell line). > > Thanks in advance > > Dave Hi, I got 2 literature citations which may be helpfull. We actually are doing these assays with rhodopsin solubilized in detergent. Read the method sections of these papers carefully, especially the concentration of detergent and also look up more stuff published by Dan Oprian. The whole issue is pretty tricky. 1) Wessling-Resnick M, Johnson, G.L. JBC 262, 3697-3705, (1987) original paper for rhodopsin aasy in ROS cells 2) Rim, J. Oprian D.D. Biochem 34, 11938-11945 rhodopsin assay with rhodopsin expressed in COS-cells good luck Carsten From owner-7tms_r@net.bio.net Tue Jun 25 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!howland.reston.ans.net!quagga.ru.ac.za!uct.ac.za!news From: david@chempath.uct.ac.za Newsgroups: bionet.molbio.proteins.7tms_r Subject: GTP gamma S binding method replies Date: Thu, 27 Jun 1996 05:42:58 GMT Organization: UCT Medical School Lines: 73 Message-ID: <4qt6og$ask@groa.uct.ac.za> NNTP-Posting-Host: med52.med.uct.ac.za X-Newsreader: Forte Agent .99b.112 Thanks to all those that responded to my request for a GTP gamma S binding method. I got a request to post all the replies as several other people are also interested in doing this assay, so here they are.... <<<<<<<<<<<< From: IJZERMAN@rulgca.LeidenUniv.nl > The assay works nice for Gi/Gq coupled receptors, not Gs. > A good reference is Lorenzen et al, Mol. Pharmacol. 44 (1993) 115- > 123. You could contact her at the University of Heidelberg, Dep. > Pharmacol., Im Neuenheimer Feld 366, 69120 Heidelberg, Germany > tel Germany-6221-548561, fax G-6221-548644 <<<<<<<<<<<< From: Carsten Schubert > I got 2 literature citations which may be helpfull. We actually are > doing these assays with rhodopsin solubilized in detergent. Read the > method sections of these papers carefully, especially the > concentration of detergent and also look up more stuff published by > Dan Oprian. The whole issue is pretty tricky. > 1) Wessling-Resnick M, Johnson, G.L. JBC 262, 3697-3705, (1987) > original paper for rhodopsin aasy in ROS cells > 2) Rim, J. Oprian D.D. Biochem 34, 11938-11945 > rhodopsin assay with rhodopsin expressed in COS-cells <<<<<<<<<<<< From: richard deth > We adapted the method of Hilf and Jacobs (Eur. J. Pharmacol. 172:155- > 163 (1989)) to PC12 cells overexpressing alpha 2D receptors. It > should be okay for COS-1 cells. Our paper was Mol. Pharm. 45:524-531 > (1994). <<<<<<<<<<<< From: krausej@thalamus.wustl.edu (Dr. Jim Krause) > One reference we found useful was that related to muscarinic > receptor stimulation of GTPgS binding and GTPase assays. Life Sci > 52, 449-456, 1993. This was CHO cells. <<<<<<<<<<<< From: marlene_jacobson@Merck.Com (Marlene Jacobson) > Lazareno and Birdsall (1993) Br. J. Pharmacol. 109:1120-1127. <<<<<<<<<<<< From: Tim Young-1 > Try this ref: Journal Receptor and Signal Transduction Res. (1995) > Vol 15 (Nos 1-4) p 199-211. > I know [35S]-GTPgS binding works well for receptors that couple to > Gi, however, I have had little success with receptors that couple > to Gs. <<<<<<<<<<<< From: "Virginia Boundy" > Try checking out references from Dana Selley in Steve Childers lab > at Bowman Grey University in North Carolina. They have used this > assay with membranes preps as well as with brain slices. I found this reference from the above group that may be of use: Proc. Natl. Acad. Sci. USA 92:7242-7246, August 1995 <<<<<<<<<<<< Thanks again to everyone Dave From owner-7tms_r@net.bio.net Wed Jun 26 23:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!swrinde!newsfeed.internetmci.com!info.ucla.edu!nnrp.info.ucla.edu!news.ucdavis.edu!bullwinkle.ucdavis.edu!not-for-mail From: szmazer@bullwinkle.ucdavis.edu (Jonathan Mazer) Newsgroups: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.yeast Subject: Re: Yeast antibodies!! Followup-To: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.yeast Date: 28 Jun 1996 00:28:23 GMT Organization: University of California, Davis Lines: 18 Message-ID: <4qv8v7$ef9@mark.ucdavis.edu> References: <4qpl38$kt3$1@mhadf.production.compuserve.com> NNTP-Posting-Host: bullwinkle.ucdavis.edu X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Xref: biosci bionet.molbio.proteins:8216 bionet.molbio.proteins.7tms_r:762 bionet.molbio.yeast:5463 DSF (76042.532@CompuServe.COM) wrote: : I'm desperately looking for commercially available antibodies to : ubiquitously expressed yeast proteins, for instance, anti-TATA : Binding Protein. HELP!! : Lynette I'm afraid you're out of luck. There are no known commercially available antibodies to yeast proteins that I know about. Your best bet is to check the literature for a lab that will have made it's own antibody(ies) against something similar to what you are working on or what you need. Write or email them and ask for it. Explain, of course, that you are not trying to copy their work with their antibody. Most labs will be willing to help. (It will take a little more time, but it should work.) Good Luck. -jsmazer@ucdavis.edu From owner-7tms_r@net.bio.net Wed Jun 26 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!newshub.csu.net!csulb.edu!not-for-mail From: lgeller@csulb.edu (Louis Geller) Newsgroups: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.yeast Subject: Re: Yeast antibodies!! Followup-To: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r,bionet.molbio.yeast Date: 28 Jun 1996 06:51:30 GMT Organization: Cal State Long Beach Lines: 15 Message-ID: <4qvvdi$4o8@hatathli.csulb.edu> References: <4qpl38$kt3$1@mhadf.production.compuserve.com> NNTP-Posting-Host: gull.csulb.edu X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Xref: biosci bionet.molbio.proteins:8219 bionet.molbio.proteins.7tms_r:764 bionet.molbio.yeast:5466 DSF (76042.532@CompuServe.COM) wrote: : I'm desperately looking for commercially available antibodies to : ubiquitously expressed yeast proteins, for instance, anti-TATA : Binding Protein. HELP!! : Lynette -- I don't know if this will help, but there is an anti-alphaCarboxypeptidase Y (CPY) antibody. This is commercially available from Molecular Probes. Louis Geller Graduate Student, Cell/Molecular Biology From owner-7tms_r@net.bio.net Wed Jun 26 23:00:00 1996 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!swrinde!newsfeed.internetmci.com!in1.uu.net!news.new-york.net!news.columbia.edu!news From: "Dr. Richard Robinson" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: program for analysis of ligand binding FOR MACINTOSH Date: Thu, 27 Jun 1996 08:33:24 -0700 Organization: Columbia University Lines: 5 Message-ID: <31D2A9C4.22A9@columbia.edu> References: NNTP-Posting-Host: ph7-w311-robinson.cpmc.columbia.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0 (Win16; I) Biosoft sells an IBM-PC version of the ligand program, and I think they offer a MAC version as well. I don't have their phone or fax number, but their address in the US is PO Box 10938, Ferguson MO, 63135. Their home office is in England (49 Bateman St, Cambridge CB2 1LR, U.K.). Hope they have what you need. From owner-7tms_r@net.bio.net Wed Jun 26 23:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!swrinde!hookup!news.nstn.ca!newsflash.concordia.ca!sunqbc.risq.net!athena.ulaval.ca!news From: jfl@rsvs.ulaval.ca (Jean-Francois Larrivee) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: BK receptor antibody search Date: Thu, 27 Jun 1996 19:16:46 GMT Organization: Centre de Recherche de l'Hotel-Dieu de Quebec Lines: 18 Message-ID: <31d2d6e3.1651594@news.ulaval.ca> References: <31D18A3D.41@ma.ultranet.com> NNTP-Posting-Host: poste21-232.crhdq.ulaval.ca X-Newsreader: Forte Agent .99d/16.182 On Wed, 26 Jun 1996 15:06:37 -0400, "John P. McGrath" wrote: >I'm in search of antibodies to the rat bradykinin B2 receptor for use >in immunohistochemical studies. >Any information re: availability of such antibodies and their use >would be greatly appreciated. > I believe that Merck has some antibodies against the mouse BK B2R (they have done immunohistochemistry on their knockout mice). Maybe they have an Ab against the rat receptor?? Good luck! Jean-Francois Larrivee From owner-7tms_r@net.bio.net Thu Jun 27 23:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!swrinde!newsfeed.internetmci.com!newsserver.jvnc.net!crick.bms.com!usenet From: John Watson Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: GTP gamma S binding assay method? Date: Fri, 28 Jun 1996 10:10:27 -0400 Organization: Bristol-Myers Squibb Company Lines: 20 Message-ID: <31D3E7D3.C9E@bms.com> References: <4qjjnu$95a@groa.uct.ac.za> Reply-To: watson_j@bms.com NNTP-Posting-Host: s107_static_223.wf.bms.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0 (Macintosh; I; PPC) Dave Myburgh wrote: > > Hi All > > Does anyone out there have a method for doing GTP gamma S binding > assays on membranes of GPCRs expressed in COS-1 cells (or any similar > cell line). You might take a look at Signal Transduction: A Practical Approach (G. Milligan, ed., IRL Press 1992), pp. 51-53. These pages present and discuss a method for doing GTPgS binding in cyc- (and other) membranes. ---------------- John Watson Bristol-Myers Squibb Co. watson_j@bms.com --------------------------------------------------------------------- "If you're not part of the solution, you're part of the precipitate." --------------------------------------------------------------------- From owner-7tms_r@net.bio.net Fri Jun 28 23:00:00 1996 Path: biosci!rutgers!uwm.edu!uwvax!newssinet!wakasato!gipwm!news From: Tokio Nakane Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: program for analysis of ligand binding FOR MACINTOSH Date: 29 Jun 1996 05:57:54 GMT Organization: Department of Pharmacology, Shinshu University School of Medicine Lines: 5 Message-ID: <4r2gl2$4bt@gipwm.shinshu-u.ac.jp> References: <31D2A9C4.22A9@columbia.edu> NNTP-Posting-Host: pharmacol3.shinshu-u.ac.jp Mime-Version: 1.0 Content-Type: text/plain; charset=iso-2022-jp Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; 68K) X-URL: news://gipwm.shinshu-u.ac.jp/31D2A9C4.22A9@columbia.edu I think you can download "macligand" from file://ftp.embl-heidelberg.de/pub/software/mac/macligand.hqx