From owner-proteins@hgmp.mrc.ac.uk Tue Nov 2 00:45:21 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id EB20F17A64; Tue, 2 Nov 1999 00:45:20 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id AAA00229 for ; Tue, 2 Nov 1999 00:45:19 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id AAA19061 for proteins-list@hgmp.mrc.ac.uk; Tue, 2 Nov 1999 00:45:18 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f Message-ID: <381E3208.2F5ABC05@yahoo.com> From: jaylino X-Mailer: Mozilla 4.5 [fr] (Win95; I) X-Accept-Language: fr MIME-Version: 1.0 X-Newsgroups: bionet.molbio.proteins Subject: Re: none References: Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit Lines: 6 Date: Tue, 02 Nov 1999 00:36:40 GMT X-Complaints-To: abuse@ulaval.ca X-Trace: carnaval.risq.qc.ca 941503000 132.203.41.113 (Mon, 01 Nov 1999 19:36:40 EST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk lili a 閏rit : Test for the same reasons From owner-proteins@hgmp.mrc.ac.uk Thu Nov 4 08:09:32 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C40B017A5A; Thu, 4 Nov 1999 08:09:31 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id IAA02376 for ; Thu, 4 Nov 1999 08:09:29 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA16447 for proteins-list@hgmp.mrc.ac.uk; Thu, 4 Nov 1999 08:09:28 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: guysjj6531@AOL.COM X-Newsgroups: bionet.molbio.proteins Subject: Confidential Information Date: 3 Nov 1999 23:48:18 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Distribution: world Message-ID: <199911040748.XAA28660@net.bio.net> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I am looking for a special person with a good work Ethic and an extraordinary desire who wants to earn $3000-$5000 per week in 30-45 days "THIS IS NOT MULTI-LEVEL MARKETING" We will give you the training and personal support you will need to ensure your success. 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NOT MLM. *FREE INFO 24 HOUR MESSAGE CALL 1-416-410-7530 rem - dfg472@excite.com From owner-proteins@hgmp.mrc.ac.uk Thu Nov 4 12:45:42 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 6480C17A6A; Thu, 4 Nov 1999 12:45:41 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id MAA23936 for ; Thu, 4 Nov 1999 12:45:15 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id MAA20020 for proteins-list@hgmp.mrc.ac.uk; Thu, 4 Nov 1999 12:45:14 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: nshh@163.NET X-Newsgroups: bionet.molbio.proteins Subject: Help:what had happened in western blot? Date: 4 Nov 1999 04:26:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Distribution: world Message-ID: <19991104201248.25382.fmail@163.net> Reply-To: nshh@163.NET To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi everyone, After the total protein of E.coli was separated by PAGE, and transferred to PDGF (and NC) membrance,a special antibody was used to do western blot as normal,but all the E.coli protein disappeared on the membrance, who can tell me what had happened? It will help me a lot. thanks David Neer ________________________________________________ 欢迎您使用163电子邮局免费服务 http://www.163.net 163超级酷全国最大的免费个人主页基地http://cool.163.net From owner-proteins@hgmp.mrc.ac.uk Thu Nov 4 13:15:14 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8163117A6A; Thu, 4 Nov 1999 13:15:13 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA25643 for ; Thu, 4 Nov 1999 13:15:11 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA20391 for proteins-list@hgmp.mrc.ac.uk; Thu, 4 Nov 1999 13:15:09 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: grima@iaf.cnrs-gif.fr (grima) X-Newsgroups: bionet.molbio.proteins Subject: antibody Date: 4 Nov 1999 13:17:28 GMT Organization: CNRS, Institut Alfred Fessard Lines: 7 Message-ID: <7vs118$1pp$2@upsn21.u-psud.fr> X-Newsreader: InterNews 2.0@mac41.iaf.cnrs-gif.fr.[U] XDisclaimer: User not authenticated To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I am looking for a commercial polyclonal antibody (rabbit) to use as standard in Drosophila western blots and which would recognize a protein of at least 100 kd in the head of drosophila. Thanks B.Grima From owner-proteins@hgmp.mrc.ac.uk Thu Nov 4 16:15:25 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8D6BD17B74; Thu, 4 Nov 1999 16:15:23 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA09997 for ; Thu, 4 Nov 1999 16:15:17 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA22753 for proteins-list@hgmp.mrc.ac.uk; Thu, 4 Nov 1999 16:15:15 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Lee Hunt X-Newsgroups: bionet.molbio.proteins Subject: Predicted vs actual MW Date: Thu, 04 Nov 1999 16:06:07 +0000 Organization: University of Sheffield, UK Lines: 4 Message-ID: <3821AEEE.9EFC3D58@sheffield.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.7 [en] (Win98; I) X-Accept-Language: en,frouzan To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Does anybody have explanations as to why a protein on a Western can differ widely from its predicted size. Other than proteolysis or removal of targetting regions what else could cause this? From owner-proteins@hgmp.mrc.ac.uk Thu Nov 4 19:05:16 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id BDF9417B57; Thu, 4 Nov 1999 19:05:15 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA21737 for ; Thu, 4 Nov 1999 19:05:14 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA24804 for proteins-list@hgmp.mrc.ac.uk; Thu, 4 Nov 1999 19:05:12 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Cornelius Krasel Subject: Re: Predicted vs actual MW X-Newsgroups: bionet.molbio.proteins References: <3821AEEE.9EFC3D58@sheffield.ac.uk> User-Agent: tin/pre-1.4-19990413 ("Endemoniada") (UNIX) (Linux/2.0.36 (i486)) Date: Thu, 4 Nov 1999 18:55:29 +0100 Message-ID: Lines: 15 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Lee Hunt wrote: > Does anybody have explanations as to why a protein on a Western can > differ widely from its predicted size. Other than proteolysis or removal > of targetting regions what else could cause this? Some proteins run irregularly on SDS-PAGE, either because they bind more or less SDS than the "average" protein, or because they do not completely unfold, even in SDS. --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP4 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@hgmp.mrc.ac.uk Fri Nov 5 01:26:14 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id BD3C817B8F; Fri, 5 Nov 1999 01:26:13 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id BAA10419; Fri, 5 Nov 1999 01:26:07 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id BAA07823; Fri, 5 Nov 1999 01:26:05 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Hoyun Lee" X-Newsgroups: bionet.molbio.proteins,bionet.general,sci.research.postdoc References: Subject: Chair in Cancer Research Lines: 47 X-Newsreader: Microsoft Outlook Express 4.72.3110.5 X-MimeOLE: Produced By Microsoft MimeOLE V4.72.3110.3 Message-ID: Date: Thu, 4 Nov 1999 20:26:36 -0500 X-Complaints-To: abuse@sprint.ca X-Trace: newscontent-01.sprint.ca 941765044 149.99.68.240 (Thu, 04 Nov 1999 20:24:04 EST) Organization: Sprint Canada Inc. To: bioforum@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Chair in Cancer Research A consortium consisting of Laurentian University (LU) the Northeastern Ontario Regional Cancer Centre (NEORCC and the H魀ital r間ional de Sudbury Regional Hospital (HRSRH) invites applications for a newly created Chair in Cancer Research. The successful applicant will have an outstanding record in an area related to the molecular biology of cancer and be recognized internationally as a leader in this field of research. The Chair will hold a senior, tenured faculty appointment in the Department of Chemistry and Biochemistry at LU and an appointment with NEORCC and Cancer Care Ontario. In addition to establishing a research program in the molecular biology of cancer, the Chair is expected to work with the members of the consortium: a) Lead the Tumour Biology group consisting of principal investigators located at NEORCC and LU. b) Assume a leadership role in the establishment of a Ph.D. program in Biomolecular Science at LU utilizing resources and expertise available within the consortium and the wider community. Plans are underway to develop an integrated research and clinical molecular biology laboratory at the HRSRH. The Chair will have an opportunity to advise the CEO of the HRSRH on the tumour biology related clinical research and development initiatives in the laboratory. The consortium is committed to equity in employment and encourages applications from all qualified candidates including women, aboriginal peoples, members of visible minorities and persons with disabilities. In accordance with Canadian immigration requirements, this advertisement is directed first to Canadian citizens and permanent residents. Appointment date is negotiable. For further information please contact: Dr. Hermann Falter, Chair Search Committee for the Chair in Cancer Research Office of the Vice President, Academic Laurentian University Sudbury, Ontario P3E 2C6 705-675-1151 ext. 3363 email: hfalter@nickel.laurentian.ca From owner-proteins@hgmp.mrc.ac.uk Fri Nov 5 10:12:57 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 4DC0E17A9B; Fri, 5 Nov 1999 10:12:57 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id KAA09359 for ; Fri, 5 Nov 1999 10:12:55 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA14281 for proteins-list@hgmp.mrc.ac.uk; Fri, 5 Nov 1999 10:12:54 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "A.F. Simpson" X-Newsgroups: bionet.molbio.proteins Subject: Re: Predicted vs actual MW Date: Fri, 05 Nov 1999 10:08:15 -0800 Organization: University of Leicester Lines: 17 Message-ID: <38231D0E.5AAE@le.ac.uk> References: <3821AEEE.9EFC3D58@sheffield.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: rook.le.ac.uk 941796444 1010791 143.210.176.54 (5 Nov 1999 10:07:24 GMT) X-Complaints-To: usenet@le.ac.uk To: Cornelius Krasel X-Mailer: Mozilla 3.01 (Win16; I) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Cornelius Krasel wrote: > Lee Hunt wrote: > > Does anybody have explanations as to why a protein on a Western can > > differ widely from its predicted size. Other than proteolysis or removal > > of targetting regions what else could cause this? > > Some proteins run irregularly on SDS-PAGE, either because they bind > more or less SDS than the "average" protein, or because they do not > completely unfold, even in SDS. Also, heavy glycosilation can cause a protein to run with an abnormally high molecular weight. > --Cornelius. love Anna From owner-proteins@hgmp.mrc.ac.uk Fri Nov 5 13:15:23 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id D0E0717A58; Fri, 5 Nov 1999 13:15:22 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA25455 for ; Fri, 5 Nov 1999 13:15:21 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA16524 for proteins-list@hgmp.mrc.ac.uk; Fri, 5 Nov 1999 13:15:19 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: suzi2@FLASHMAIL.COM X-Newsgroups: bionet.molbio.proteins Subject: We Will Pay Your Bills Date: 5 Nov 1999 05:02:13 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 107 Distribution: world Message-ID: <00000f63610b$0000011b$000007b1@mail.debis.dk> Reply-To: sela24@FLASHMAIL.COM To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk ><><><><><><><><><><><><><><><><><><><><> "Opt-In" <><><><><><><><><><><><><><><><><><><><><> To Unsubscribe your name and email from our Opt-In listserver, see end of message. 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From owner-proteins@hgmp.mrc.ac.uk Fri Nov 5 13:18:15 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id F1D2517A58; Fri, 5 Nov 1999 13:18:14 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA25857 for ; Fri, 5 Nov 1999 13:18:12 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA16649 for proteins-list@hgmp.mrc.ac.uk; Fri, 5 Nov 1999 13:18:11 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: sela24@flashmail.com X-Newsgroups: bionet.molbio.proteins Subject: We Will Pay Your Bills Date: 5 Nov 1999 13:18:10 -0000 Organization: MRC Human Genome Mapping Project Resource Centre Lines: 108 Message-ID: <00005364557b$00002d22$000007ab@mailgate.ilhadamagia.com.br> X-Trace: niobium.hgmp.mrc.ac.uk 941807890 16646 193.62.192.80 (5 Nov 1999 13:18:10 GMT) X-Complaints-To: news@hgmp.mrc.ac.uk X-Received: from mserv3.dl.ac.uk (IDENT:root@mserv3.dl.ac.uk [148.79.80.28]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA25840; Fri, 5 Nov 1999 13:18:06 GMT X-Received: from nt-server.itl.co.il ([209.88.190.98]) by mserv3.dl.ac.uk (8.9.3/8.9.3/[ref postmaster@dl.ac.uk]) with ESMTP id NAA07678; Fri, 5 Nov 1999 13:18:20 GMT X-Received: from mailgate.ilhadamagia.com.br ([63.20.221.128]) by nt-server.itl.co.il (Netscape Mail Server v2.02) with SMTP id AAT420; Fri, 5 Nov 1999 15:22:07 +0200 X-To: X-Priority: 3 X-MSMail-Priority: Normal X-Reply-To: sela24@flashmail.com To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk ><><><><><><><><><><><><><><><><><><><><> "Opt-In" <><><><><><><><><><><><><><><><><><><><><> To Unsubscribe your name and email from our Opt-In listserver, see end of message. Dear Subscriber, We can Slash your bills,from 40-80% off of what you owe.That's money that goes back into your pocket! _WHAT WE DO Negotiate with your creditors to SAVE you money! Relieve you from creditor phone calls. We can work on all types of debt,for example: CREDIT CARDS VENDORS SUPPLIERS LAWSUITS JUDGEMENTS MEDICAL BILLS INSURANCE BILLS LOANS _COST NO RESULTS-NO FEE! We work on a contingency basis,our fee is based on a percentage of the savings-if we can't help you SAVE money,there is NO FEE! Our company has been saving individuals and businesses money for over 18 years - DON'T HESITATE - call our toll free number NOW for more information. 1-888-900-0125 to get STARTED ! ****************************************************************** If you have recieved this message in error please email sela24@flashmail.com julesb@flashmail.com suzi2@flashmail.com to unsubscribe in the subject field. --- From owner-proteins@hgmp.mrc.ac.uk Fri Nov 5 16:45:32 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 2E80017A82; Fri, 5 Nov 1999 16:45:30 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA14700 for ; Fri, 5 Nov 1999 16:45:28 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA19195 for proteins-list@hgmp.mrc.ac.uk; Fri, 5 Nov 1999 16:45:27 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: rogier Subject: Re: Help:what had happened in western blot? X-Newsgroups: bionet.molbio.proteins Message-ID: <2750ac20.e9f61ab5@usw-ex0102-013.remarq.com> Lines: 14 Bytes: 649 X-Originating-Host: 130.37.84.156 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <19991104201248.25382.fmail@163.net> X-Wren-Trace: ePrf9/bvqOKpqfrn+L/p0+vs5vP997Xg8PHst+bg6vfz7rv1/7z65uK1+vq067q9taWhu7Xx1MvPzA== Date: Fri, 05 Nov 1999 08:28:10 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 941819465 10.0.2.13 (Fri, 05 Nov 1999 08:31:05 PST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk how do you know your protein was on the membrane? did you stain with ponceau or amido black or something like that? if it was there, it should stay on. if you blocked your membrane with protein (BSA, milk, ...), you have no way to find out if your e. coli protein is there, cos there'll be protein all over the membrane. maybe your antibody doesn't work. try reprobing your blot with another antibody to see if you pick up a signal. if you do, it means your blot is OK. Good luck, Rogier * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Sat Nov 6 10:02:26 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 242C617A4E; Sat, 6 Nov 1999 10:02:25 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id KAA05160; Sat, 6 Nov 1999 10:02:24 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA10167; Sat, 6 Nov 1999 10:02:23 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Per Mygind X-Newsgroups: bionet.microbiology,bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: [Fwd: Detection of Protein glycosylation] Date: Sat, 06 Nov 1999 10:53:02 +0100 Organization: University of Aarhus, Department of Computer Science (DAIMI) Lines: 95 Message-ID: <3823FA7E.9A5AF1CE@biobase.dk> Mime-Version: 1.0 Content-Type: multipart/mixed; boundary="------------E7B0FF3F77B1C6C3A6DE62CE" X-Trace: xinwen.daimi.au.dk 941881964 8844560 255.255.255.255 (6 Nov 1999 09:52:44 GMT) X-Complaints-To: news@daimi.au.dk X-Mailer: Mozilla 4.07 [en] (X11; I; SunOS 5.7 sun4u) To: methods@hgmp.mrc.ac.uk, microbio@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk This is a multi-part message in MIME format. --------------E7B0FF3F77B1C6C3A6DE62CE Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit -- Venlig Hilsen Per Mygind ************************************************************************ If you are are not part of the solution, you are part of the precipitate ************************************************************************ Per Mygind, Cand.scient Department of Medical Microbiology and Immunology The Bartholin Building, University of Aarhus, Denmark phone : 89 42 17 47, fax : 86 19 61 28 ************************************************************************ It's hard work and great art to make life not so serious ************************************************************************ Life is what happens to you while you're busy making other plans (John Lennon) All you touch and all you see is all your life will ever be (Roger Waters) --------------E7B0FF3F77B1C6C3A6DE62CE Content-Type: message/rfc822 Content-Transfer-Encoding: 7bit Content-Disposition: inline Path: news.daimi.au.dk!not-for-mail From: Per Mygind Newsgroups: bionet.glycosci Subject: Detection of Protein glycosylation Date: Sat, 06 Nov 1999 10:51:22 +0100 Organization: University of Aarhus, Department of Computer Science (DAIMI) Message-ID: <3823FA1A.E5CF74CF@biobase.dk> NNTP-Posting-Host: mg04.gram.au.dk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: xinwen.daimi.au.dk 941881864 8844560 255.255.255.255 (6 Nov 1999 09:51:04 GMT) X-Complaints-To: news@daimi.au.dk NNTP-Posting-Date: 6 Nov 1999 09:51:04 GMT X-Mailer: Mozilla 4.07 [en] (X11; I; SunOS 5.7 sun4u) Xref: news.daimi.au.dk bionet.glycosci:1169 Dear Readers I',m working with a Bacteria and I've reasons to suspect that some proteins in the outer membrane are glycosylated. What is the best way to detect such protein modification, is there any more sensitive way than using Schiff's reagent and to digest the sample with heparinase. I'm not sure what kind of glycosylation is involved. Any help is greatly appreciated -- Yours sincerely Per Mygind ************************************************************************ If you are are not part of the solution, you are part of the precipitate ************************************************************************ Per Mygind, Cand.scient Department of Medical Microbiology and Immunology The Bartholin Building, University of Aarhus, Denmark phone : 89 42 17 47, fax : 86 19 61 28 ************************************************************************ It's hard work and great art to make life not so serious ************************************************************************ Life is what happens to you while you're busy making other plans (John Lennon) All you touch and all you see is all your life will ever be (Roger Waters) --------------E7B0FF3F77B1C6C3A6DE62CE-- From owner-proteins@hgmp.mrc.ac.uk Sat Nov 6 11:40:48 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 30AA417A4E; Sat, 6 Nov 1999 11:40:48 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id LAA07745 for ; Sat, 6 Nov 1999 11:40:45 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id LAA11258 for proteins-list@hgmp.mrc.ac.uk; Sat, 6 Nov 1999 11:40:45 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: wuafiorue41@ejf.hu (39yeh474) X-Newsgroups: bionet.molbio.proteins Subject: Find out ANYTHING about ANYONE! Date: 6 Nov 1999 03:45:04 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 50 Distribution: world Message-ID: <199911061603AAA16079@3893hook3hd3.rand.fr> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk ******************************************************* This list is opt-in only. We are linked to many web sites that offer free subscriptions to our opt-in list. You will be removed from this list at any time by following the simple instructions that can be found at the end of this email. THIS IS NOT SPAM! You are on our mailing list because you subscribed or someone you know subscribed for you at one of our associate web sites. ******************************************************* Cyber Investigator "EASY WAY TO FIND OUT ANYTHING ABOUT ANYONE" http://www1.50megs.com/bert99/ Cyber Investigator TAKES YOU BEYOND WHAT SEARCH ENGINES CAN DO! 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You may rcmove your name from our opt-in list by sending an e-mail to the address below with the word rcmove i From owner-proteins@hgmp.mrc.ac.uk Sat Nov 6 15:49:24 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 457F917A4E; Sat, 6 Nov 1999 15:49:24 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id PAA14198 for ; Sat, 6 Nov 1999 15:49:22 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA14224 for proteins-list@hgmp.mrc.ac.uk; Sat, 6 Nov 1999 15:49:21 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: pxpst2+@pitt.edu (Peter) X-Newsgroups: bionet.molbio.proteins Subject: Re: Predicted vs actual MW Date: Sat, 06 Nov 1999 10:43:13 -0400 Organization: University Of Pittsburgh Lines: 14 Message-ID: References: <3821AEEE.9EFC3D58@sheffield.ac.uk> Mail-Copies-To: pxpst2+@pitt.edu X-Newsreader: MT-NewsWatcher 2.4.4 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article <3821AEEE.9EFC3D58@sheffield.ac.uk>, Lee Hunt wrote: > Does anybody have explanations as to why a protein on a Western can > differ widely from its predicted size. Other than proteolysis or removal > of targetting regions what else could cause this? Always remember that PAGE does not seperate purely by mass/size. Seperation is by the charge to mass (m/z). Alter this and and you will have abherent migration. Regards, Peter Pediaditakis From owner-proteins@hgmp.mrc.ac.uk Sat Nov 6 22:25:17 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id D618917A43; Sat, 6 Nov 1999 22:25:16 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA20013 for ; Sat, 6 Nov 1999 22:25:15 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA27109 for proteins-list@hgmp.mrc.ac.uk; Sat, 6 Nov 1999 22:25:14 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Marcin Wierzbicki X-Newsgroups: bionet.molbio.proteins Subject: Ubiquitin Date: Sat, 6 Nov 1999 23:20:06 +0100 Organization: Academic Computer Centre CYFRONET - Krakow Lines: 6 Message-ID: Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, Please give me any information about ubiquitin. I need it for my thesis. Thank you. From owner-proteins@hgmp.mrc.ac.uk Sun Nov 7 00:45:25 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 2B44017A43; Sun, 7 Nov 1999 00:45:23 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id AAA02495; Sun, 7 Nov 1999 00:45:21 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id AAA28810; Sun, 7 Nov 1999 00:45:21 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Hoyun Lee" X-Newsgroups: bionet.molbio.proteins,bionet.general,sci.research.postdoc References: Subject: Chair in Cancer Research Lines: 49 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 Message-ID: Date: Sat, 6 Nov 1999 19:46:43 -0500 X-Complaints-To: abuse@sprint.ca X-Trace: newscontent-01.sprint.ca 941935448 149.99.67.97 (Sat, 06 Nov 1999 19:44:08 EST) Organization: Sprint Canada Inc. To: bioforum@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Chair in Cancer Research A consortium consisting of Laurentian University (LU) the Northeastern Ontario Regional Cancer Centre (NEORCC and the H魀ital r間ional de Sudbury Regional Hospital (HRSRH) invites applications for a newly created Chair in Cancer Research. The successful applicant will have an outstanding record in an area related to the molecular biology of cancer and be recognized internationally as a leader in this field of research. The Chair will hold a senior, tenured faculty appointment in the Department of Chemistry and Biochemistry at LU and an appointment with NEORCC and Cancer Care Ontario. In addition to establishing a research program in the molecular biology of cancer, the Chair is expected to work with the members of the consortium: a) Lead the Tumour Biology group consisting of principal investigators located at NEORCC and LU. b) Assume a leadership role in the establishment of a Ph.D. program in Biomolecular Science at LU utilizing resources and expertise available within the consortium and the wider community. Plans are underway to develop an integrated research and clinical molecular biology laboratory at the HRSRH. The Chair will have an opportunity to advise the CEO of the HRSRH on the tumour biology related clinical research and development initiatives in the laboratory. The consortium is committed to equity in employment and encourages applications from all qualified candidates including women, aboriginal peoples, members of visible minorities and persons with disabilities. In accordance with Canadian immigration requirements, this advertisement is directed first to Canadian citizens and permanent residents. Appointment date is negotiable. For further information please contact: Dr. Hermann Falter, Chair Search Committee for the Chair in Cancer Research Office of the Vice President, Academic Laurentian University Sudbury, Ontario P3E 2C6 705-675-1151 ext. 3363 email: hfalter@nickel.laurentian.ca From owner-proteins@hgmp.mrc.ac.uk Sun Nov 7 15:45:55 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1530F17A43; Sun, 7 Nov 1999 15:45:54 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id PAA29215 for ; Sun, 7 Nov 1999 15:45:53 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA09313 for proteins-list@hgmp.mrc.ac.uk; Sun, 7 Nov 1999 15:45:52 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: pxpst2+@pitt.edu (Peter) X-Newsgroups: bionet.molbio.proteins Subject: Re: Ubiquitin Date: Sun, 07 Nov 1999 10:21:46 -0400 Organization: University Of Pittsburgh Lines: 10 Message-ID: References: Mail-Copies-To: pxpst2+@pitt.edu X-Newsreader: MT-NewsWatcher 2.4.4 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article , Marcin Wierzbicki wrote: > Please give me any information about ubiquitin. > I need it for my thesis. Used to tag proteins for distruction by proteosome. Look up on public med the term ubiquination. Peter From owner-proteins@hgmp.mrc.ac.uk Mon Nov 8 04:06:21 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1490D17A81; Mon, 8 Nov 1999 04:06:20 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id EAA22867 for ; Mon, 8 Nov 1999 04:06:19 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id EAA26442 for proteins-list@hgmp.mrc.ac.uk; Mon, 8 Nov 1999 04:06:18 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Nick Theodorakis Subject: Re: Predicted vs actual MW X-Newsgroups: bionet.molbio.proteins Message-ID: <0a0133f8.4e007621@usw-ex0101-002.remarq.com> Lines: 32 Bytes: 1266 X-Originating-Host: 63.15.102.90 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <3821AEEE.9EFC3D58@sheffield.ac.uk> X-Wren-Trace: eAovBwYfWBJZRBcBB0UHAgsmDgcKAU4THQcOABA+BlgIDwQZC0kXARUfCx1KTx0IS1BVS0wAJDs6OFgzOg== Date: Sun, 07 Nov 1999 19:58:25 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon1 942033871 10.0.2.2 (Sun, 07 Nov 1999 20:04:31 PST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article , pxpst2+@pitt.edu (Peter) wrote: > In article <3821AEEE.9EFC3D58@sheffield.ac.uk>, Lee Hunt > wrote: > > Does anybody have explanations as to why a protein on a Western > can > > differ widely from its predicted size. Other than proteolysis or > removal > > of targetting regions what else could cause this? > Always remember that PAGE does not seperate purely by mass/size. > Seperation is by the charge to mass (m/z). Alter this and and you > will > have abherent migration. > Regards, > Peter Pediaditakis That's the textbook version for "pure" electrophoresis, anyway. In real life, (for biochemists/molecular biologists, anyway), molecular sieving is important. Otherwise, all DNA should have the same electrophoretic mobility, and to a first approximation, all SDS-covered proteins should too. Actually, all DNA does have the same m/z and have the same electrophoretic mobility and would run the same place, except that those pesky agarose or polyacrylamide strands get in the way. Nick * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Mon Nov 8 13:10:17 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 0B06E17A61; Mon, 8 Nov 1999 13:10:16 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA25635 for ; Mon, 8 Nov 1999 13:10:15 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA03146 for proteins-list@hgmp.mrc.ac.uk; Mon, 8 Nov 1999 13:10:13 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: mmerckel@btk.utu.fi (Michael Merckel) X-Newsgroups: bionet.molbio.proteins Subject: 1kD concentrator ? Date: 8 Nov 1999 13:10:12 -0000 Organization: MRC Human Genome Mapping Project Resource Centre Lines: 11 Message-ID: <3826CBAF.6BCBDB0D@btk.utu.fi> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 942066613 3143 193.62.192.80 (8 Nov 1999 13:10:13 GMT) X-Complaints-To: news@hgmp.mrc.ac.uk X-Received: from post.it.helsinki.fi (post-ether.it.helsinki.fi [128.214.4.122]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA25619 for ; Mon, 8 Nov 1999 13:10:09 GMT X-Received: from btk.utu.fi (ala.biocenter.helsinki.fi [128.214.88.36]) by post.it.helsinki.fi (8.9.3/8.9.3-SPAMmers-sod-off) with ESMTP id PAA08167 for ; Mon, 8 Nov 1999 15:10:08 +0200 (EET) X-Mailer: Mozilla 4.51C-SGI [en] (X11; I; IRIX 6.5 IP32) X-Accept-Language: en X-To: proteins@hgmp.mrc.ac.uk To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, I am interested in finding a 1kD cutoff centricon type concentrator for small volumes, <100ul end volume. Thanks for any tips. Mike --- From owner-proteins@hgmp.mrc.ac.uk Mon Nov 8 17:10:49 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8865A17A8A; Mon, 8 Nov 1999 17:10:48 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id RAA14781 for ; Mon, 8 Nov 1999 17:10:46 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA06212 for proteins-list@hgmp.mrc.ac.uk; Mon, 8 Nov 1999 17:10:45 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Jan Aelen X-Newsgroups: bionet.molbio.proteins Subject: Re: 1kD concentrator ? Date: Mon, 08 Nov 1999 16:48:17 +0100 Organization: University of Nijmegen, The Netherlands Lines: 19 Message-ID: <3826F0BF.F3AA6631@nmr.kun.nl> References: <3826CBAF.6BCBDB0D@btk.utu.fi> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.06 [en] (Win95; I) To: Michael Merckel To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Maybe an idea: www.spectrumlabs.com Michael Merckel wrote: > Hi, > > I am interested in finding a 1kD cutoff centricon type > concentrator for small volumes, <100ul end volume. > Thanks for any tips. > > Mike > > --- -- J.Aelen e-mail: Janal@nmr.kun.nl From owner-proteins@hgmp.mrc.ac.uk Mon Nov 8 17:17:58 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 9FCFE17A8A; Mon, 8 Nov 1999 17:17:57 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id RAA15560 for ; Mon, 8 Nov 1999 17:17:55 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA06362 for proteins-list@hgmp.mrc.ac.uk; Mon, 8 Nov 1999 17:17:54 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: genacad@aol.com (Genacad) X-Newsgroups: bionet.molbio.proteins Subject: SPSS SigmaStat 5.0 for Win9x/NT $478 Lines: 57 X-Admin: news@aol.com Date: 08 Nov 1999 15:56:29 GMT Organization: AOL http://www.aol.com Message-ID: <19991108105629.20987.00001643@ng-fi1.aol.com> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk We offer a complete range of statistics and graphing software from SPSS for Students, Faculty and Staff of recognized educational institutions. 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For more information, or to place an order, e-mail -- or contact -- Genesis Technologies Inc Toll Free : 800-433-6326 Fax : 814-231-0371 E-Mail : sales@genesis-technologies.com Web Site : http://www.genesis-technologies.com ============================================================= From owner-proteins@hgmp.mrc.ac.uk Mon Nov 8 19:39:42 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 7D89317A60; Mon, 8 Nov 1999 19:39:41 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA22735 for ; Mon, 8 Nov 1999 19:39:39 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA08066 for proteins-list@hgmp.mrc.ac.uk; Mon, 8 Nov 1999 19:39:38 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Gabriel Birrane X-Newsgroups: bionet.molbio.proteins Subject: Protein solubility; Sarcosyl and Triton Date: Mon, 08 Nov 1999 14:38:25 -0800 Organization: Harvard Institutes of Medicine Lines: 9 Message-ID: <382750E1.E1FCE232@yahoo.com> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: plato.harvard.edu 942089677 9660 12.6.221.19 (8 Nov 1999 19:34:37 GMT) X-Complaints-To: usenet@harvard.edu X-Mailer: Mozilla 4.51C-SGI [en] (X11; I; IRIX64 6.5 IP28) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I have a protein that's soluble in sarcosyl but becomes insoluble after the addition of triton. Is there an alternative to triton that I can use to negate the effects of sarcosyl while keeping my protein soluble? Any suggestions are appreciated. -Judy From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 04:51:34 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B917117A43; Tue, 9 Nov 1999 04:51:33 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id EAA14466; Tue, 9 Nov 1999 04:51:31 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id EAA22967; Tue, 9 Nov 1999 04:51:29 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Hein Subject: Re: [Fwd: Detection of Protein glycosylation] X-Newsgroups: bionet.molbio.proteins,bionet.microbiology,bionet.molbio.methds-reagnts Message-ID: <0a0133f8.525a9830@usw-ex0107-049.remarq.com> Lines: 8 Bytes: 274 X-Originating-Host: 63.23.252.193 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <3823FA7E.9A5AF1CE@biobase.dk> X-Wren-Trace: eAksBAUcWxFaQRgIAhtQIAIbEQMDQ0oQAUVQWU5PQwdUWhtGVhBP Date: Mon, 08 Nov 1999 20:41:39 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 942122674 10.0.2.49 (Mon, 08 Nov 1999 20:44:34 PST) To: methods@hgmp.mrc.ac.uk, microbio@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Try anti-carbohydrate Westerns, or Westerns with Abs against your proteins before and after EndoF / EndoH digestion. * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 05:00:23 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 280DC17AA3; Tue, 9 Nov 1999 05:00:22 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id FAA14722 for ; Tue, 9 Nov 1999 05:00:21 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id FAA23083 for proteins-list@hgmp.mrc.ac.uk; Tue, 9 Nov 1999 05:00:16 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Hein Subject: Re: Need help:insoluble to soluble X-Newsgroups: bionet.molbio.proteins Message-ID: <028ffd81.54cd3ce0@usw-ex0107-049.remarq.com> Lines: 9 Bytes: 355 X-Originating-Host: 63.23.252.193 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: X-Wren-Trace: eD4bMzIrbCZtdi8/NSxnFzUsJjQ0dH0nNnJnbnl4dDBjbSxxYSd4 Date: Mon, 08 Nov 1999 20:51:02 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 942123238 10.0.2.49 (Mon, 08 Nov 1999 20:53:58 PST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Try purifying the inclusion body, solubilizing it in 8M urea, and slowly refolding it. Works wonders for many proteins; esp. if you have an affinity column to help select active refolded proteins. * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 08:02:58 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 2E4A417A43; Tue, 9 Nov 1999 08:02:57 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id IAA25856; Tue, 9 Nov 1999 08:02:56 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA25295; Tue, 9 Nov 1999 08:02:55 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Hein Subject: Re: [Fwd: Detection of Protein glycosylation] X-Newsgroups: bionet.molbio.proteins,bionet.microbiology,bionet.molbio.methds-reagnts Message-ID: <31171c80.511656ad@usw-ex0107-049.remarq.com> Lines: 8 Bytes: 272 X-Originating-Host: 63.23.252.193 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <3823FA7E.9A5AF1CE@biobase.dk> X-Wren-Trace: eOTB6ejxtvy3rPXl7/a9ze/2/O7urqf97Ki9tKOiruq5t/aru/2i Date: Mon, 08 Nov 1999 20:36:48 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 942122501 10.0.2.49 (Mon, 08 Nov 1999 20:41:41 PST) To: methods@hgmp.mrc.ac.uk, microbio@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Try anti-carbohydrate Westerns, or Westerns with Ab's against your proteins before and after EndoF/EndoH treatment * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 10:21:19 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 33F0417A61; Tue, 9 Nov 1999 10:21:18 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id KAA05048 for ; Tue, 9 Nov 1999 10:21:16 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA27130 for proteins-list@hgmp.mrc.ac.uk; Tue, 9 Nov 1999 10:21:13 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "PAUL MACLEAN" X-Newsgroups: bionet.molbio.proteins Subject: Small Mr Standards Date: Tue, 9 Nov 1999 00:09:42 -0500 Organization: Prodigy Internet http://www.prodigy.com Lines: 11 Message-ID: <808a49$4d9q$1@newssvr03-int.news.prodigy.com> X-Trace: newssvr03-int.news.prodigy.com 942123977 3970311 209.252.88.198 (9 Nov 1999 05:06:17 GMT) X-Complaints-To: abuse@prodigy.net X-Newsreader: Microsoft Outlook Express 4.72.3155.0 X-Mimeole: Produced By Microsoft MimeOLE V4.72.3155.0 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Is anyone familiar will commercially available, small molecular weight standards (500-1000 Daltons) that could be used in the second dimension of 2D-gel electrophoresis? If not, is it feasible to make some standards myself? Any ideas or direction would be appreciated. Paul MacLean ECU SOM From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 10:21:23 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 095BE17A61; Tue, 9 Nov 1999 10:21:22 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id KAA05047 for ; Tue, 9 Nov 1999 10:21:16 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA27134 for proteins-list@hgmp.mrc.ac.uk; Tue, 9 Nov 1999 10:21:14 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f Message-ID: <3827B0B9.20A97AC7@indo.net.id> From: suhendar X-Mailer: Mozilla 4.7 [en] (Win98; I) X-Accept-Language: en MIME-Version: 1.0 X-Newsgroups: bionet.molbio.proteins Subject: test Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 2 Date: Tue, 09 Nov 1999 12:27:21 +0700 X-Trace: news 942151141 202.154.43.89 (Tue, 09 Nov 1999 05:39:01 GMT) Organization: RADnet To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk test transmisi From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 14:28:10 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 08E3517A59; Tue, 9 Nov 1999 14:28:09 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id OAA25405; Tue, 9 Nov 1999 14:28:08 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA00324; Tue, 9 Nov 1999 14:28:07 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Michael Brunsteiner X-Newsgroups: sci.bio.technology,sci.med,sci.bio.misc,sci.bio.food-science,bionet.molbio.proteins,bionet.microbiology Subject: looking for molec-bio mailing-lists Date: Tue, 09 Nov 1999 15:24:56 +0100 Organization: Vienna University, Austria Lines: 15 Message-ID: <38282EB8.C7D72338@mdy.univie.ac.at> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: www.univie.ac.at 942157496 46992 131.130.40.10 (9 Nov 1999 14:24:56 GMT) X-Complaints-To: news-adm@news.univie.ac.at X-Mailer: Mozilla 4.05 [en] (X11; U; IRIX 5.3 IP22) To: microbio@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I'm looking for mailing-lists on molecular biology, biotechnology and related fields (actually offers for PhD positions) any recommendations ? thanx for hints! mic _____________________________________________________ Michael Brunsteiner From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 16:46:36 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C409117A43; Tue, 9 Nov 1999 16:46:35 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA06391; Tue, 9 Nov 1999 16:46:32 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA02937; Tue, 9 Nov 1999 16:46:31 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Donald D. Bustamante" X-Newsgroups: sci.bio.technology,sci.med,sci.bio.misc,sci.bio.food-science,bionet.molbio.proteins,bionet.microbiology Subject: Re: looking for molec-bio mailing-lists Date: Tue, 09 Nov 1999 09:36:00 -0700 Organization: National Biotechnology Information Facility Lines: 30 Message-ID: <38284D70.C2F@nbif.org> References: <38282EB8.C7D72338@mdy.univie.ac.at> Reply-To: dbustama@nbif.org Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: bubba.NMSU.Edu 942164965 11042 128.123.36.225 (9 Nov 1999 16:29:25 GMT) X-Complaints-To: usenet@bubba.NMSU.Edu X-Mailer: Mozilla 3.01Gold (WinNT; I) To: microbio@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Visit the National Biotechnology Information Facility WWW site for a compilation of these lists http://www.nbif.org Michael Brunsteiner wrote: > > I'm looking for mailing-lists on > molecular biology, biotechnology > and related fields > (actually offers for PhD positions) > any recommendations ? > > thanx for hints! > > mic > > > _____________________________________________________ > Michael Brunsteiner > > -- Donald D. Bustamante, Project Manager/Co-PI National Biotechnology Information Facility Box 30002, Dept. 3548 Las Cruces, New Mexico USA 88003 (505)-646-5031 FAX: (505)-522-9373 URL: http://www.nbif.org From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 19:35:27 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id AD77D17A78; Tue, 9 Nov 1999 19:35:25 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA16508 for ; Tue, 9 Nov 1999 19:35:21 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA04932 for proteins-list@hgmp.mrc.ac.uk; Tue, 9 Nov 1999 19:35:18 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Marcel Dinger" X-Newsgroups: bionet.molbio.proteins Subject: Genamics SoftwareSeek Date: Wed, 10 Nov 1999 08:28:57 +1300 Organization: The Internet Group (Hamilton) Lines: 21 Message-ID: <942175893.217874@ham.ihug.co.nz> X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2615.200 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 Cache-Post-Path: ham.ihug.co.nz!unknown@p57-max7.ham.ihug.co.nz X-Cache: nntpcache 2.3.3 (see http://www.nntpcache.org/) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk GENAMICS SOFTWARESEEK The Largest Science Software Directory on Earth. http://genamics.com/software/ Genamics SoftwareSeek now indexes well over 1000 freely-distributable and commercial titles. Classifications include DNA sequence analysis, protein structure analysis, RNA structure prediction, molecular modeling, image analysis, laboratory utilities, and sequence alignment. All major platforms are supported, including, Windows, Macintosh, Linix, and Unux. Over 150 online tools and resources that run directly through your internet browser are also listed. New software can be submitted online through our web-site. Marcel Dinger, Genamics. http://genamics.com/software/ From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 19:45:22 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 6F21C17A78; Tue, 9 Nov 1999 19:45:21 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA17007 for ; Tue, 9 Nov 1999 19:45:19 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA05060 for proteins-list@hgmp.mrc.ac.uk; Tue, 9 Nov 1999 19:45:17 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Koen Peeters X-Newsgroups: bionet.molbio.proteins Subject: to differentiate between 2 proteins Date: Tue, 09 Nov 1999 20:31:46 +0100 Organization: University of Ghent, Belgium Lines: 26 Message-ID: <382876A2.24026AF6@gengenp.rug.ac.be> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.5 [en] (Win95; I) X-Accept-Language: en Cc: anpic@gengenp.rug.ac.be To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Dear, I need a solution to the following problem : I have a Fab fragment with and without C-terminal DIKDEL aminoacids produced in plants. I need to know wether the C-terminal DIKDEL are present or cleaved in planta. Is there any molecular technique that I could apply that will differentiate the presence of these 6 C-terminal aminoacids? No antibodies against DIKDEL/KDEL are working on Western blot. Thanks Koen -- ================================================================== Koen Peeters DEPARTMENT OF GENETICS Fax:32 (0)9 2645349 UNIVERSITY OF GENT, K. L. Ledeganckstraat 35, B-9000 Gent, Belgium Vlaams Instituut voor Biotechnologie VIB mailto:kopee@gengenp.rug.ac.be http://www.plantgenetics.rug.ac.be ================================================================== From owner-proteins@hgmp.mrc.ac.uk Tue Nov 9 22:45:52 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B43F617A59; Tue, 9 Nov 1999 22:45:51 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA28718; Tue, 9 Nov 1999 22:45:49 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA15467; Tue, 9 Nov 1999 22:45:48 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Anne Thomas" X-Newsgroups: sci.bio.technology,sci.med,sci.bio.misc,sci.bio.food-science,bionet.molbio.proteins,bionet.microbiology Subject: Re: looking for molec-bio mailing-lists Date: Tue, 9 Nov 1999 22:34:52 -0000 Organization: NTL Internet News Service Lines: 24 Message-ID: <80a7pn$u1$1@nclient7-gui.server.ntli.net> References: <38282EB8.C7D72338@mdy.univie.ac.at> X-Trace: nclient7-gui.server.ntli.net 942187127 961 212.49.254.166 (9 Nov 1999 22:38:47 GMT) X-Complaints-To: abuse@net.ntl.com X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 To: microbio@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Theres another newsgroup that you dont have on your list. bionet.molbio.methds-reagnts Anne :o) Michael Brunsteiner wrote in message news:38282EB8.C7D72338@mdy.univie.ac.at... > I'm looking for mailing-lists on > molecular biology, biotechnology > and related fields > (actually offers for PhD positions) > any recommendations ? > > thanx for hints! > > mic > > > _____________________________________________________ > Michael Brunsteiner > > From owner-proteins@hgmp.mrc.ac.uk Wed Nov 10 01:06:16 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 2149517AA9; Wed, 10 Nov 1999 01:06:14 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id BAA02679 for ; Wed, 10 Nov 1999 01:06:12 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id BAA17149 for proteins-list@hgmp.mrc.ac.uk; Wed, 10 Nov 1999 01:06:10 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Hein Subject: Re: protein binding to immobilised ligand X-Newsgroups: bionet.molbio.proteins Message-ID: <070dbf92.912e9ec0@usw-ex0106-044.remarq.com> Lines: 9 Bytes: 384 X-Originating-Host: 63.75.53.102 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <7stdvb$7ic_001@leeds.ac.uk> X-Wren-Trace: eOvO5uf+ufO4o/rq4PmywuD58+Hhoajy46eyu6yop+WxvuW7tfk= Date: Tue, 09 Nov 1999 16:52:25 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 942195320 10.0.2.44 (Tue, 09 Nov 1999 16:55:20 PST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk .. also, the immobilization of the ligand may affect the binding affinity due to chemical modification of the ligand itself. i.e. the type of linking may affect the amount of free ligand you need to use to elute your protein. * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Wed Nov 10 01:15:33 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 972E517AA9; Wed, 10 Nov 1999 01:15:31 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id BAA02949 for ; Wed, 10 Nov 1999 01:15:28 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id BAA17269 for proteins-list@hgmp.mrc.ac.uk; Wed, 10 Nov 1999 01:15:26 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Hein Subject: Re: quantitating protein in solid sample X-Newsgroups: bionet.molbio.proteins Message-ID: <065f8722.93003882@usw-ex0106-044.remarq.com> Lines: 10 Bytes: 396 X-Originating-Host: 63.75.53.102 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <000501bf02ed$60c39b80$a87debc2@nieuwenhuizen> X-Wren-Trace: eI+qgoOa3Zfcx56OhJ3WpoSdl4WFxcyWh8PW38jMw4HV2oHf0Z0= Date: Tue, 09 Nov 1999 16:59:23 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 942195739 10.0.2.44 (Tue, 09 Nov 1999 17:02:19 PST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk 8 M urea or 6 M guanidine hydrochloride will efficiently extract your proteins from the solid. To keep it solid and analyse it, nitrogen content is routinely used in food chemistry. Dissolve it and you can use a number of assay methods. * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Wed Nov 10 16:41:32 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 7929F17A67; Wed, 10 Nov 1999 16:41:32 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA29649 for ; Wed, 10 Nov 1999 16:41:30 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA28558 for proteins-list@hgmp.mrc.ac.uk; Wed, 10 Nov 1999 16:41:29 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Lutz Thon X-Newsgroups: bionet.molbio.proteins Subject: yeast-protein-extraction and following identification of a nuclear protein Date: Wed, 10 Nov 1999 17:34:21 +0100 Organization: Rechenzentrum der Universitaet Kiel, Germany Lines: 60 Message-ID: <38299E8D.ABA0EF43@mail.uni-kiel.d400.de> Mime-Version: 1.0 Content-Type: multipart/mixed; boundary="------------239FB01CE292D9F64CE15DD9" X-Trace: infosrv.rz.uni-kiel.de 942251517 29735 134.245.169.12 (10 Nov 1999 16:31:57 GMT) X-Complaints-To: usenet@infosrv.rz.uni-kiel.de X-Mailer: Mozilla 4.06 [en] (Win95; I) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk This is a multi-part message in MIME format. --------------239FB01CE292D9F64CE15DD9 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit I am trying to detect a nuclear protein from Sacc. cerevisiae by a western-blot. I used a glass-bead + vortex method to disrupt the cells in the following buffer: (glass bead disruption buffer) 20 mM Tris-Cl, pH 7.9 10 mM MgCl2 1 mM EDTA 5%(v/v) Glycerol 1 mM DTT 0.3 M ammonium sulfate no protease inhibitors After disruption (checked by microscope) I spinned down the debris, and used the supernatant for SDS-PAGE and blotting. No signal was recieved from this extraction. But when i treated the debris from above with urea buffer (8 M urea, 100 mM Na2HPO4, 10 mM Tris-Cl, pH 8), centrifuged, an used this supernatant for western-blotting i got a weak signal. Questions: Why is my target protein not in the first supernatant? Is the nucleus from yeast cells destroyed by the glass-bead + vortex method? (my target protein is in the nucleus) My target protein is a DNA-binding protein, is it stripped off from DNA by the glass-bead disruption buffer mentioned above? If not, what buffer would be suitable? Do you know a better method for small-scale protein extraction from yeast (especially for a DNA-binding protein)? Thanks, Lutz --------------239FB01CE292D9F64CE15DD9 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Lutz Thon Content-Disposition: attachment; filename="vcard.vcf" begin: vcard fn: Lutz Thon n: Thon;Lutz org: University of Kiel, Germany email;internet: stu32394@mail.uni-kiel.d400.de x-mozilla-cpt: ;0 x-mozilla-html: FALSE version: 2.1 end: vcard --------------239FB01CE292D9F64CE15DD9-- From owner-proteins@hgmp.mrc.ac.uk Wed Nov 10 18:05:16 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3FA2217A43; Wed, 10 Nov 1999 18:05:16 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id SAA05363 for ; Wed, 10 Nov 1999 18:05:14 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id SAA29511 for proteins-list@hgmp.mrc.ac.uk; Wed, 10 Nov 1999 18:05:13 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Frank Fuerst X-Newsgroups: bionet.molbio.proteins Subject: Re: to differentiate between 2 proteins Date: Wed, 10 Nov 1999 18:57:23 +0100 Lines: 14 Message-ID: <80cbhs$pu3g$1@fu-berlin.de> References: <382876A2.24026AF6@gengenp.rug.ac.be> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: fu-berlin.de 942256508 850032 141.89.207.60 (16 49 955) X-Mailer: Mozilla 4.51 [de]C-CCK-MCD QXW03200 (WinNT; I) X-Accept-Language: de,en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Koen Peeters wrote: > > > Is there any molecular technique that I could apply that will > differentiate the presence > of these 6 C-terminal aminoacids? A good MALDI-TOF mass spectrometer should be able to resolve the difference, or perhaps other MS-techniques. Good luck, Frank Fuerst -- Hi! I'm Norton Antivirus. Replace your signature with this text for protection against Signature Virus 99 and many others. From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 09:51:45 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id ADA7017ABD; Thu, 11 Nov 1999 09:51:44 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id JAA18676 for ; Thu, 11 Nov 1999 09:51:41 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id JAA19011 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 09:51:40 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Byung-Hoon Kim X-Newsgroups: bionet.molbio.proteins Subject: Re: yeast-protein-extraction and following identification of a nuclear protein Date: Thu, 11 Nov 1999 10:49:18 +0100 Organization: Zentrum fuer Datumverarbeitung Lines: 56 Message-ID: <382A911D.D4D954B4@uni-tuebingen.de> References: <38299E8D.ABA0EF43@mail.uni-kiel.d400.de> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: newsserv.zdv.uni-tuebingen.de 942313685 1640 134.2.88.165 (11 Nov 1999 09:48:05 GMT) X-Complaints-To: usenet@newsserv.uni-tuebingen.de X-Mailer: Mozilla 4.5 [de] (WinNT; I) X-Accept-Language: de,ko To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Lutz Thon schrieb: > I am trying to detect a nuclear protein from Sacc. cerevisiae by a > western-blot. > I used a glass-bead + vortex method to disrupt the cells in the > following buffer: > (glass bead disruption buffer) > 20 mM Tris-Cl, pH 7.9 > 10 mM MgCl2 > 1 mM EDTA > 5%(v/v) Glycerol > 1 mM DTT > 0.3 M ammonium sulfate > no protease inhibitors > > After disruption (checked by microscope) I spinned down the debris, > and used the supernatant for SDS-PAGE and blotting. > No signal was recieved from this extraction. > > But when i treated the debris from above with urea buffer (8 M urea, 100 > > mM Na2HPO4, 10 mM Tris-Cl, pH 8), centrifuged, an used this supernatant > for western-blotting i got a weak signal. > > Questions: > Why is my target protein not in the first supernatant? > Is the nucleus from yeast cells destroyed by the glass-bead + vortex > method? (my target protein is in the nucleus) > My target protein is a DNA-binding protein, is it stripped off from DNA > > by the glass-bead disruption buffer mentioned above? > If not, what buffer would be suitable? > Do you know a better method for small-scale protein extraction from > yeast (especially for a DNA-binding protein)? > > Thanks, Lutz Hello Lutz, I could detect an yeast transcription factor (which binds constitutively to DNA) using glass beads and the following lysis buffer. 1 X PBS containing 1mM EDTA, 5% Glycerol, 0.1% NP-40 and protease inhibitor. The isolated protein could be succesfully used for band shift assay. I'm not sure whether the 'NP-40' is an essential component, but it may help you. Byung-Hoon Zentrum fuer MolekularBiologie der Pflanzen Uni Tuebingen From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 12:42:25 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 256C617A63; Thu, 11 Nov 1999 12:42:25 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id MAA01321 for ; Thu, 11 Nov 1999 12:42:19 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id MAA21155 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 12:42:18 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Jan Aelen X-Newsgroups: bionet.molbio.proteins Subject: Impact system Date: Thu, 11 Nov 1999 13:32:35 +0100 Organization: University of Nijmegen, The Netherlands Lines: 15 Message-ID: <382AB762.92F805BA@nmr.kun.nl> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.06 [en] (Win95; I) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi all, Can anyone tell me your experience about the Impact-Cn System of "New England Biolabs" as alternative to the conventional method for recombinant protein expression to express a target protein as a fusion to an affinity tag protein such as GST. thanks in advance -- Jan Aelen e-mail: Janal@nmr.kun.nl From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 18:25:13 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C355917B2D; Thu, 11 Nov 1999 18:25:12 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id SAA28167 for ; Thu, 11 Nov 1999 18:25:10 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id SAA25486 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 18:25:09 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Kresten X-Newsgroups: bionet.molbio.proteins Subject: Re: Predicted vs actual MW Date: Thu, 11 Nov 1999 18:11:54 GMT Organization: Deja.com - Before you buy. Lines: 34 Message-ID: <80f0t5$5ae$1@nnrp1.deja.com> References: <3821AEEE.9EFC3D58@sheffield.ac.uk> X-Article-Creation-Date: Thu Nov 11 18:11:54 1999 GMT X-Http-User-Agent: Mozilla/4.5 [en] (Win95; I) X-Http-Proxy: 1.0 www2.crc.dk:3128 (Squid/2.1.RELEASE), 1.0 x41.deja.com:80 (Squid/1.1.22) for client 130.226.182.217, 130.226.183.6 X-MyDeja-Info: XMYDJUIDkresten To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article , pxpst2+@pitt.edu (Peter) wrote: > In article <3821AEEE.9EFC3D58@sheffield.ac.uk>, Lee Hunt > wrote: [snip] > Always remember that PAGE does not seperate purely by mass/size. > Seperation is by the charge to mass (m/z). Alter this and and you > will have abherent migration. This is why we include SDS in our PAGEs. If it were so that proteins bound a number of SDS molecules proportinal to their mass and that this number was so large that the proteins own charge would be negligable compared to the charges that SDS bring, then all proteins in SDS would have approximately the same m/z. The idea is then, that you separate on m (or rather volume or some other structure parameter) using the sieving effect of the PA gel. Kresten > > Regards, > Peter Pediaditakis > -- The address kresten@my-dejanews.com is for spambots only. Please mail me at LysLeuLeu@crc.dk transforming the pre@-part into my initials. Kresten Lindorff Larsen, Dept. Yeast Genetics Sent via Deja.com http://www.deja.com/ Before you buy. From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 19:33:33 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8648717B22; Thu, 11 Nov 1999 19:33:32 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA01561 for ; Thu, 11 Nov 1999 19:33:27 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA26371 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 19:33:25 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: ding2@is09.fas.harvard.edu (-Hua Ding) X-Newsgroups: bionet.molbio.proteins Subject: Protein Expression and Purification Position at Pfizer Cambridge Site Date: 11 Nov 1999 19:07:03 GMT Organization: Harvard University, Cambridge, MA Lines: 13 Message-ID: <80f44n$uhv$1@news.fas.harvard.edu> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk How to Respond: Please note: Job code BN110-DYH is required for all communications regarding this position at Pfizer Discovery Technology Center. Via Email openings@cambridge.pfizer.com Via Fax 617-551-3111 From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 19:35:12 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 234AA17B22; Thu, 11 Nov 1999 19:35:11 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA01787 for ; Thu, 11 Nov 1999 19:35:10 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA26379 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 19:35:09 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: ding2@is09.fas.harvard.edu (-Hua Ding) X-Newsgroups: bionet.molbio.proteins Subject: Protein Expression and Purification Position at Pfizer Cambridge Site Date: 11 Nov 1999 19:00:33 GMT Organization: Harvard University, Cambridge, MA Lines: 13 Message-ID: <80f3oh$u1v$1@news.fas.harvard.edu> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk How to Respond: Please note: Job code BN110-DYH is required for all communications regarding this position at Pfizer Discovery Technology Center. Via Email openings@cambridge.pfizer.com Via Fax 617-551-3111 From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 20:20:56 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 66BFC17A4E; Thu, 11 Nov 1999 20:20:56 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id UAA04056 for ; Thu, 11 Nov 1999 20:20:54 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id UAA26996 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 20:20:53 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Schmidt.Thorsten@gmx.de X-Newsgroups: bionet.molbio.proteins Subject: Bacteria contamination in yeast two hybrid screening Date: Thu, 11 Nov 1999 20:12:58 GMT Organization: Deja.com - Before you buy. Lines: 63 Message-ID: <80f806$b00$1@nnrp1.deja.com> X-Article-Creation-Date: Thu Nov 11 20:12:58 1999 GMT X-Http-User-Agent: Mozilla/4.0 (compatible; MSIE 5.0; Windows 98; DigExt) X-Http-Proxy: 1.1 x39.deja.com:80 (Squid/1.1.22) for client 166.124.48.202 X-MyDeja-Info: XMYDJUIDschmidt_thorsten To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, I have a serious problem and hope that you could help me: I am conducting a yeast two hybrid screen using the GAL 4-system and the Y190 yeast strain). My problem is that almost all of my plates (80 !) of my yeast screening seem to be contaminated with bacteria!!! The plates are now incubating for 6 days and several yeast cultures (approx. 0.5 mm diameter)have formed all over the plates, as expected. But all over the plates (in the area not covered by yeast colonies)a light "smear" is visible. I streaked over a part of some plates and streaked the cells on a slide. Under the microscope many healthy yeast are visible as well as some yeast with clear bacteria IN it (they are moving in the yeasts cells) as well as single bacteria cells and some clusters of bacteria forming around lysed yeast cells. After that,I picked single yeast colonies, they are looking healthy and hardly unaffected. What can I do now? Do I have to throw all these 80 (!) plates away and start a new screening? Or could I still use them? Will the bacteria seriously affected the results of the screening? Could I somehow "spray" any antibiotic on the plates? Could I (to prevent this problem in future screenings) just add antibiotics (Pen/Strep or something else) to the yeast SC-media plates? If yes at which concentration? Will the antibiotic affect the yeast or the results of the screening? Normally, I would like to incubate the plates for another four days before performing the beta-Galactosidase-Assay. Should I wait these days or should I proceed as fast as possible? I would imagine that the E.coli might have overtaken the complete plates and killed all yeasts if I will wait longer. Do you have experiences with that? Thank you so much in advance for your answer! Everything would help! Thorsten Schmidt Sent via Deja.com http://www.deja.com/ Before you buy. From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 20:33:04 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 6B81D17A4E; Thu, 11 Nov 1999 20:33:03 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id UAA04464 for ; Thu, 11 Nov 1999 20:33:01 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id UAA27120 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 20:32:55 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: ding2@is08.fas.harvard.edu (-Hua Ding) X-Newsgroups: bionet.molbio.proteins Subject: Protein Expression and Purification Position at Pfizer Cambridge Site Date: 11 Nov 1999 20:25:44 GMT Organization: Harvard University, Cambridge, MA Lines: 29 Message-ID: <80f8o8$ujs$1@news.fas.harvard.edu> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Work in a unique environment to develop cutting edge, protein-based and cell-based assay systems for high throughput drug discoveries. Work to include recombinant DNA and cell culture techniques; protein expression (in bacteria, insect and mammalian cell lines), purification and characterization (HPLC, FPLC and fluorescent spectroscopy etc.); involve in the development of new biochemical and biophysical platforms to characterize protein-ligand interactions. Computer experience, strong cell culture skills, good communication skills and evidence of effective team interaction are required. Two to three years experience is desirable. B.S. in chemistry or biological sciences is required and M.S. is preferred. How to Respond: Please note: Job code BN110-DYH is required for all communications regarding this position at Pfizer Discovery Technology Center. Via Email: openings@cambridge.pfizer.com Via Fax: 617-551-311 From owner-proteins@hgmp.mrc.ac.uk Thu Nov 11 22:45:18 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B355617B64; Thu, 11 Nov 1999 22:45:17 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA13690 for ; Thu, 11 Nov 1999 22:45:12 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA07014 for proteins-list@hgmp.mrc.ac.uk; Thu, 11 Nov 1999 22:45:10 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Huseyin Kucuktas X-Newsgroups: bionet.molbio.proteins Subject: Creatine Kinase Date: Thu, 11 Nov 1999 16:22:17 -0600 Organization: Auburn University Usenet Server Lines: 16 Message-ID: References: <80f8o8$ujs$1@news.fas.harvard.edu> Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII X-Trace: ultranews.duc.auburn.edu 942358340 19778 131.204.2.23 (11 Nov 1999 22:12:20 GMT) X-Complaints-To: swangdb@ultranews.duc.auburn.edu X-Sender: kucukhu@mallard In-Reply-To: <80f8o8$ujs$1@news.fas.harvard.edu> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Dear collegues, We are trying to look at the creatine kinase enzyme activitiy of different tissues of channel catfish. We run starch gels and histochemically looked at the enzyme activities by using postcoupling technique. We used acidic and basic buffers and run total proteins. We got high enzyme activity at cardiac, skeletal and smooth (stomach) muscles. However, in both acidic and basic buffer conditions, the stomach bands showed up at the cathodal side. Does anybody have any clue about cathodal CK enzymes? Thanks. Huseyin Kucuktas From owner-proteins@hgmp.mrc.ac.uk Fri Nov 12 03:40:17 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 92C8317A8E; Fri, 12 Nov 1999 03:40:16 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id DAA20859 for ; Fri, 12 Nov 1999 03:40:14 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id DAA10633 for proteins-list@hgmp.mrc.ac.uk; Fri, 12 Nov 1999 03:40:13 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: fvilla@BUCARAMANGA.CETCOL.NET.CO ("FEDERICO VILLALOBOS R") X-Newsgroups: bionet.molbio.proteins Subject: Antropogenetics Date: 11 Nov 1999 19:33:26 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Distribution: world Message-ID: To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Dear Dr: I an working with human population in Santander-Colombia-S.A. I am interested in people who can work with us. My web is www.antropogenetica.4mg.com. Sincerily, Federico Villalobos Antropogenetics Deparment of Biological Sciences Universidad Industrial de Santander E-mail: fvilla@b-manga.cetcol.net.co fvilla40@hotmail.com Web: http://www.antropogenetica.4mg.com From owner-proteins@hgmp.mrc.ac.uk Fri Nov 12 15:35:10 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id DA5B517A5B; Fri, 12 Nov 1999 15:35:06 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id PAA02981 for ; Fri, 12 Nov 1999 15:35:02 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA19752 for proteins-list@hgmp.mrc.ac.uk; Fri, 12 Nov 1999 15:35:01 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: mmerckel@btk.utu.fi (Michael Merckel) X-Newsgroups: bionet.molbio.proteins Subject: Poly Glu in E.coli Date: 12 Nov 1999 15:34:59 -0000 Organization: MRC Human Genome Mapping Project Resource Centre Lines: 12 Message-ID: <382C339A.4B1976E5@btk.utu.fi> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 942420900 19750 193.62.192.80 (12 Nov 1999 15:35:00 GMT) X-Complaints-To: news@hgmp.mrc.ac.uk X-Received: from post.it.helsinki.fi (post-ether.it.helsinki.fi [128.214.4.122]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id PAA02974 for ; Fri, 12 Nov 1999 15:34:54 GMT X-Received: from btk.utu.fi (ala.biocenter.helsinki.fi [128.214.88.36]) by post.it.helsinki.fi (8.9.3/8.9.3-SPAMmers-sod-off) with ESMTP id RAA00723 for ; Fri, 12 Nov 1999 17:34:53 +0200 (EET) X-Mailer: Mozilla 4.51C-SGI [en] (X11; I; IRIX 6.5 IP32) X-Accept-Language: en X-To: proteins@hgmp.mrc.ac.uk To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hello, I'd appreciate any comments on trying to express a protein construct with 12 consecutive glutamates in E.coli. Anyone have any experiences with such strectches in E.coli ? Thanks Mike --- From owner-proteins@hgmp.mrc.ac.uk Fri Nov 12 16:25:22 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3ADC617AB4; Fri, 12 Nov 1999 16:25:20 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA06520 for ; Fri, 12 Nov 1999 16:25:18 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA20506 for proteins-list@hgmp.mrc.ac.uk; Fri, 12 Nov 1999 16:25:16 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f Date: Fri, 12 Nov 1999 17:21:18 +0100 From: stagljar@vetbio.unizh.ch (Igor Stagljar) X-Newsgroups: bionet.molbio.proteins Subject: Contract yeast two-hybrid technology Message-ID: X-Newsreader: MT-NewsWatcher 2.4.4 X-Original-NNTP-Posting-Host: 130.60.120.47 X-Trace: 12 Nov 1999 17:22:19 +0100, 130.60.120.47 Organization: University of Zurich Lines: 69 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Introduction Interactions between proteins regulate nearly every cellular process. The most powerful in vivo method in the identification of protein-protein interactions is the yeast two-hybrid system and variations thereof. Scientists working in different fields encounter yeast as a model system very often for the first time. Newcomers are therefore puzzled and experience difficulties when they try to establish specific yeast system in their laboratories. This procedure can be very time consuming and also cost intensive. We provide a solution: DUALSYSTEMS Our aim We are a team of molecular biologists and biochemists with a long experience in yeast hybrid screenings. We provide a yeast one- and two- hybrid based technology platform for functional characterization of protein-protein interactions. We offer high quality and fast screenings at competitive prices. Our customers will receive - a detail written report on the screening - a DNA sample of the bait dependent clones - DNA sequences of positive clones (package dependent) - map of interaction domain (package dependent) Legal notice - DUALSYSTEMS guarantees strict confidentiality - results are interpreted to the best of our knowledge - results are provided within 4 months after receiving the order - if there are no positive clones resulting from a screening, the customer pays only 60% of the package price - DUALSYSTEMS reserves to change the price of the service offered at any time Contact address: DUALSYSTEMS Institute of Veterinary Biochemistry University of Zurich Winterthurerstr. 190 CH-8057 Zurich Tel: +41-1-635 54 74 Fax: +41-1-635 68 40 Email: dualsys@vetbio.unizh.ch -- _____________________________________________________________________ Dr. Igor Stagljar Institut fur Veterinarbiochemie Tel: (41-1)-635-54-79 Universitat Zurich-Irchel Fax: (41-1)-635-68-16 Winterthurerstrasse 190 CH-8057 Zurich Switzerland _____________________________________________________________________ From owner-proteins@hgmp.mrc.ac.uk Fri Nov 12 22:33:53 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3F15E17A76; Fri, 12 Nov 1999 22:33:53 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA27313 for ; Fri, 12 Nov 1999 22:33:51 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA03544 for proteins-list@hgmp.mrc.ac.uk; Fri, 12 Nov 1999 22:33:50 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: linkman X-Newsgroups: bionet.molbio.proteins Subject: receptor identification Date: Fri, 12 Nov 1999 14:32:53 -0800 Organization: ITServices, University of British Columbia Lines: 17 Message-ID: <382C957A.23C0B37F@home.com> Reply-To: linkman1@home.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: nntp.itservices.ubc.ca 942445905 29796 137.82.151.51 (12 Nov 1999 22:31:45 GMT) X-Complaints-To: abuse@interchange.ubc.ca X-Mailer: Mozilla 4.6 (Macintosh; I; PPC) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk We are looking for a postdoctoral fellow/scientist with experience in identifying the unknown receptor for a small molecule. This position is for an industry-university project with a potential to be a permanent job in the local pharmaceutical company. If you are interested, please call 604-875-5530 or email to wjia@unixg.ubc.ca. Please note that you must have extensive experience in protein purification and ligand/receptor interaction assay. Thank for your attention. William Jia Associate Professor University of British Columbia Vancouver, Canada From owner-proteins@hgmp.mrc.ac.uk Sat Nov 13 10:12:05 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 4503C17A5B; Sat, 13 Nov 1999 10:12:04 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id KAA20579 for ; Sat, 13 Nov 1999 10:12:02 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA11670 for proteins-list@hgmp.mrc.ac.uk; Sat, 13 Nov 1999 10:12:00 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: biohelp@hgmp.mrc.ac.uk (BIOSCI Administrator) X-Newsgroups: bionet.molbio.proteins Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 13 Nov 1999 02:00:14 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 239 Distribution: world Message-ID: <199911131000.CAA01717@net.bio.net> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk (LAST REVISION: 14-AUG-99) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- All BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- the UK-HGMP-Resource Centre (known as hgmp.mrc.ac.uk): ----------------------------------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@hgmp.mrc.ac.uk. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to majordomo@hgmp.mrc.ac.uk. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Please ask for help at biosci@hgmp.mrc.ac.uk if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. From owner-proteins@hgmp.mrc.ac.uk Mon Nov 15 21:15:47 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 59C3517ACA; Mon, 15 Nov 1999 21:15:45 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id VAA08305 for ; Mon, 15 Nov 1999 21:15:38 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id VAA11429 for proteins-list@hgmp.mrc.ac.uk; Mon, 15 Nov 1999 21:15:35 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: gilchrie@SODA.DENT.UNC.EDU ("Eric P. Gilchrist") X-Newsgroups: bionet.molbio.proteins Subject: FS: Electroporator Date: 15 Nov 1999 12:54:34 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Distribution: world Message-ID: <38307197.3B3E319C@soda.dent.unc.edu> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk For Sale to any investigator: BTX 810-T Square-wave pulse electorporator w/Cuvette Chamber and 100mm Tissue Culture dish electrode. For information contact: Eric Gilchrist Univ. North Carolina School of Dentistry 919.966.2746 From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 01:44:18 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 2744317A58; Tue, 16 Nov 1999 01:44:17 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id BAA23166 for ; Tue, 16 Nov 1999 01:44:16 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id BAA21971 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 01:44:11 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: klenchin@REMOVE_TO_REPLY.facstaff.wisc.edu (Dima Klenchin) X-Newsgroups: bionet.molbio.proteins Subject: Re: Predicted vs actual MW Date: Tue, 16 Nov 1999 01:35:59 GMT Organization: UW Lines: 33 Message-ID: <80qcea$p0e$1@news.doit.wisc.edu> References: <3821AEEE.9EFC3D58@sheffield.ac.uk> <80f0t5$5ae$1@nnrp1.deja.com> X-Newsreader: News Xpress 2.01 X-No-Archive: yes To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article <80f0t5$5ae$1@nnrp1.deja.com>, Kresten wrote: :In article , : pxpst2+@pitt.edu (Peter) wrote: :> In article <3821AEEE.9EFC3D58@sheffield.ac.uk>, Lee Hunt :> wrote: :[snip] : :> Always remember that PAGE does not seperate purely by mass/size. :> Seperation is by the charge to mass (m/z). Alter this and and you :> will have abherent migration. : :This is why we include SDS in our PAGEs. If it were so that proteins :bound a number of SDS molecules proportinal to their mass and that this :number was so large that the proteins own charge would be negligable :compared to the charges that SDS bring, then all proteins in SDS would :have approximately the same m/z. The idea is then, that you separate on :m (or rather volume or some other structure parameter) using the sieving :effect of the PA gel. All is right but it's worth remembering that nothing is ideal under the moon. In case of SDS PAGE, the number of SDS molecules bound per MW is _not_ the same for all sequences. Thus, the mobility. The bottom line is that SDS PAGE as a criterion for estimation of MW is better than gel filtration but can still be very far from true MW based on sequence - even when corrected by known posttranslational modifications. Some examples: phosphorylation (32+3x18 Da) at single site can upshift the band by as much as ca 5 kDa, calmodulin runs 18 or 22 KDa depending on the presence of Ca++, myristoylation of ARF1 upshifts the band on a gel, but myristoylation of ARF6 (95% homology, but a very different pI) results in downshift. Dima From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 12:15:25 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id A354817A62; Tue, 16 Nov 1999 12:15:24 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id MAA25262 for ; Tue, 16 Nov 1999 12:15:22 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id MAA29817 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 12:15:21 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: zequabue63@KOWON.DONGSEO.AC.KR (kiowp) X-Newsgroups: bionet.molbio.proteins Subject: 'Net small cap newsletter Date: 16 Nov 1999 04:06:18 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Distribution: world Message-ID: <199911162059WAA19856@mquweo.norwich.co.nz> To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Internet & small cap stock... newsletter... f-r-e-e! We are currently offering at no charge an internet and small cap newsletter FREE for a limited time. This is a weekly newsletter that is short, to the point and will only take a couple of minutes to read. If you do not enjoy this newsletter you can unsubscribe at any time, no obligation. You have nothing to lose by subscribing, so give it a try ! ! ! To subscribe, email froth45@bigfoot.com with the subject capnews To be removed, email froth45@bigfoot.com with the subject delete thanks...thanks...thanks... From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 13:25:20 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1757817A62; Tue, 16 Nov 1999 13:25:19 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA29671; Tue, 16 Nov 1999 13:25:12 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA00689; Tue, 16 Nov 1999 13:25:09 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: tschantz@acpub.duke.edu (William Tschantz) X-Newsgroups: bionet.molbio.proteins,bionet.cellbiol Subject: H2O2 assay Date: 16 Nov 1999 13:19:21 GMT Organization: Duke University, Durham, NC, USA Lines: 17 Message-ID: <80rlkp$jfs$1@news.duke.edu> X-Newsreader: trn 4.0-test67 (15 July 1998) To: cellbiol@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi I am looking for a sensitive assay for H2O2 (hydrogen peroxide). I think the enzyme that i am working on may generate it. I am thinking of possible using a peroxidase linked assay. Has anyone done this before? Any help would be appreciated! Thanks in advance Bill -- Bill Tschantz, Ph.D. Duke University Medical Center Dept of Pharmacology and Cancer Biology C307 Levine Science Research Center (LSRC) From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 13:45:15 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5478D17A62; Tue, 16 Nov 1999 13:45:13 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA04276 for ; Tue, 16 Nov 1999 13:45:09 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA00934 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 13:45:07 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Ina Hinners X-Newsgroups: bionet.molbio.proteins Subject: Re: Predicted vs actual MW Date: Tue, 16 Nov 1999 13:34:39 +0100 Organization: Imperial Cancer Research Fund Lines: 11 Message-ID: <38314F4E.427C@icrf.icnet.uk> References: <3821AEEE.9EFC3D58@sheffield.ac.uk> <80f0t5$5ae$1@nnrp1.deja.com> <80qcea$p0e$1@news.doit.wisc.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0 (Macintosh; I; PPC) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I dont know about ARF6, but ARF1 shifts definitely to lower MW upon myristoylation. -- Ina Hinners ICRF Secretory Pathways Laboratory 44 Lincolns Inn Fields WC2A 3PX London UK email: I.Hinners@icrf.icnet.uk From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 15:25:18 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id D8F9817ABA; Tue, 16 Nov 1999 15:25:17 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id PAA15529 for ; Tue, 16 Nov 1999 15:25:13 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA03009 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 15:25:11 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Byung-Hoon Kim X-Newsgroups: bionet.molbio.proteins Subject: Can proteinase inhibitors penetrate the cell membrane? Date: Tue, 16 Nov 1999 16:18:00 +0100 Organization: Zentrum fuer Datenverarbeitung Lines: 14 Message-ID: <383175A8.3865DDA3@uni-tuebingen.de> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: newsserv.zdv.uni-tuebingen.de 942765279 6758 134.2.88.147 (16 Nov 1999 15:14:39 GMT) X-Complaints-To: usenet@newsserv.uni-tuebingen.de X-Mailer: Mozilla 4.6 [de] (WinNT; I) X-Accept-Language: de To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, all Can proteinase inhibitors (Aprotinin, Leupeptin and Pepstatin) penetrate the cell membrane? Byung-Hoon Kim ----------------------------------- Center for Plant Molecular Biology Univ. Tuebingnen Germany ----------------------------------- From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 17:15:34 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 9665717AC3; Tue, 16 Nov 1999 17:15:33 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id RAA26214 for ; Tue, 16 Nov 1999 17:15:26 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA04531 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 17:15:25 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Fergus.Doherty@nottingham.ac.uk (Fergus Doherty) X-Newsgroups: bionet.molbio.proteins Subject: Re: Can proteinase inhibitors penetrate the cell membrane? Date: Tue, 16 Nov 1999 17:05:30 +0000 Organization: Nottingham University Lines: 25 Message-ID: References: <383175A8.3865DDA3@uni-tuebingen.de> Mime-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Trace: oyez.ccc.nottingham.ac.uk 942771969 23751 128.243.183.228 (16 Nov 1999 17:06:09 GMT) X-Complaints-To: usenet@news.nottingham.ac.uk X-Newsreader: Yet Another NewsWatcher 2.4.0 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article <383175A8.3865DDA3@uni-tuebingen.de>, Byung-Hoon Kim wrote: > Hi, all > > Can proteinase inhibitors (Aprotinin, Leupeptin and Pepstatin) penetrate > the cell membrane? > > > Byung-Hoon Kim No, leupeptin can't, not sure about the others but I doubt it. They are taken up by endocytosis and act on lysosomal proteases AFAIK. However, there are some protease inhibitors which reputedly are membrane permeant. The one I know is E64-d, a dervative of the cysteine protease inhibitor E64 (which is membrane impermeant). E64-d will inhibit cytoplasmic calpain I believe. -- Fergus Doherty, School of Biomedical Sciences, Nottingham University, Fergus.Doherty@nottingham.ac.uk 0115 970 9366 (74-41366 internal) From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 17:45:27 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 003FF17B03; Tue, 16 Nov 1999 17:45:25 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id RAA28154 for ; Tue, 16 Nov 1999 17:45:19 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA04888 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 17:45:15 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: klenchin@facstaff.REMOVE_TO_REPLY.wisc.edu (Dima Klenchin) X-Newsgroups: bionet.molbio.proteins Subject: Re: Can proteinase inhibitors penetrate the cell membrane? Date: Tue, 16 Nov 1999 17:35:51 GMT Organization: UW-Madison Lines: 10 Message-ID: <80s4h9$qrg$1@news.doit.wisc.edu> References: <383175A8.3865DDA3@uni-tuebingen.de> Mime-Version: 1.0 Content-Type: text/plain; charset=KOI8-R Content-Transfer-Encoding: 8bit X-Newsreader: News Xpress 2.01 X-No-Archive: yes To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk : :Hi, all : :Can proteinase inhibitors (Aprotinin, Leupeptin and Pepstatin) penetrate :the cell membrane? Aprotinin and Leupeptin cannot for sure. Pepstatin, being a hydrophobic peptide, probably can, but probably not very efficiently. - Dima From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 21:03:36 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3B7B617B30; Tue, 16 Nov 1999 21:03:36 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id VAA09855 for ; Tue, 16 Nov 1999 21:03:34 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id VAA07437 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 21:03:33 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Cornelius Krasel Subject: Re: Can proteinase inhibitors penetrate the cell membrane? X-Newsgroups: bionet.molbio.proteins References: <383175A8.3865DDA3@uni-tuebingen.de> User-Agent: tin/pre-1.4-19990413 ("Endemoniada") (UNIX) (Linux/2.0.36 (i486)) Date: Tue, 16 Nov 1999 20:28:48 +0100 Message-ID: Lines: 14 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Byung-Hoon Kim wrote: > Can proteinase inhibitors (Aprotinin, Leupeptin and Pepstatin) penetrate > the cell membrane? Leupeptin can be used to inhibit proteolysis in lysosomes (I have seen several publications demonstrating this), but I think that it reaches them by endocytosis, not through permeation of the plasma membrane. --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP4 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 22:55:22 1999 Return-Path: Delivered-To: proteins-archive@hgmp.mrc.ac.uk Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 0B05B17A4E; Tue, 16 Nov 1999 22:55:21 +0000 (GMT) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA20066 for ; Tue, 16 Nov 1999 22:55:20 GMT Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA17029 for proteins-list@hgmp.mrc.ac.uk; Tue, 16 Nov 1999 22:55:15 GMT X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Nick Theodorakis Subject: Re: Can proteinase inhibitors penetrate the cell membrane? X-Newsgroups: bionet.molbio.proteins Message-ID: <1415c574.a42a44e5@usw-ex0102-011.remarq.com> Lines: 17 Bytes: 585 X-Originating-Host: 128.151.74.69 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here References: <383175A8.3865DDA3@uni-tuebingen.de> X-Wren-Trace: eBUwGBkARw1GWwgeGFoYHRQ5ERgVHlEMAhgRHw8hGUcXEBsGFFYIHgoAFAJVUAIXVEhLQkwbICQ7Il07Iyw= Date: Tue, 16 Nov 1999 14:38:06 -0800 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 942792693 10.0.2.11 (Tue, 16 Nov 1999 14:51:33 PST) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article <383175A8.3865DDA3@uni-tuebingen.de>, Byung-Hoon Kim wrote: > Hi, all > Can proteinase inhibitors (Aprotinin, Leupeptin and Pepstatin) > penetrate > the cell membrane? Have you thought about using small, non-peptide inhibitors like PMSF or TPCK? I'm not sure if either of these will penetrate the cell membrane, but they may be more likely to than peptide inhibitors. Nick * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@hgmp.mrc.ac.uk Tue Nov 16 23:55:29 1999 Return-Path: Delivered-To: