From owner-repertoires@net.bio.net Wed Nov 05 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Chris Jeffers Newsgroups: bionet.molecules.repertoires Subject: single chain antibody libraries Date: 6 Nov 1997 11:02:52 -0800 Organization: University of Houston Lines: 11 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <34554F31.41C6@dna.bchs.uh.edu> NNTP-Posting-Host: net.bio.net Does anyone know if there are any companies which still market randomized single chain antibody libraries (Such as those used for phage display). I would really appreciate it. Chris Jeffers University of Houston cjeffers@dna.bchs.uh.edu -- From owner-repertoires@net.bio.net Wed Nov 05 22:00:00 1997 Path: biosci!biosci!not-for-mail From: jose luis toran Newsgroups: bionet.molecules.repertoires Subject: Pcomb3H vector Date: 6 Nov 1997 11:02:47 -0800 Organization: cnb Lines: 6 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <344FCB57.5A94@cnb.uam.es> Reply-To: jltora@cnb.uam.es NNTP-Posting-Host: net.bio.net hello i'm trying to make library using the pcomb3H system anyone know if is there any place i can get the whole sequences in file? From owner-repertoires@net.bio.net Wed Nov 05 22:00:00 1997 Path: biosci!biosci!not-for-mail From: SODECM Newsgroups: bionet.molecules.repertoires Subject: Human cDNA library from adipose tisse Date: 6 Nov 1997 11:03:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 42 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <01bce659$528a3b80$a5b7a8c6@bumhead> NNTP-Posting-Host: net.bio.net ------=_NextPart_000_0004_01BCE62F.69B43380 Content-type: text/plain; charset="iso-8859-1" Content-transfer-encoding: quoted-printable Hello, I have searched the catalouges and internet for a Human cDNA expression = library to no avail. Does anyone know of where I could obtain such a = library? Ian Murray imurra@po-box.mcgill.ca ------=_NextPart_000_0004_01BCE62F.69B43380 Content-type: text/html; charset="iso-8859-1" Content-transfer-encoding: quoted-printable

Hello,

I have searched the catalouges and = internet for=20 a Human cDNA expression library to no avail. Does anyone know of where I = could=20 obtain such a library?

Ian Murray

imurra@po-box.mcgill.ca

------=_NextPart_000_0004_01BCE62F.69B43380-- From owner-repertoires@net.bio.net Wed Nov 05 22:00:00 1997 Path: biosci!biosci!not-for-mail From: John Newsgroups: bionet.molecules.repertoires Subject: Compound Library Date: 6 Nov 1997 11:03:03 -0800 Organization: AsInEx ltd. Lines: 10 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <34562ABB.439A@online.ru> NNTP-Posting-Host: net.bio.net As of October 1997 the inventory at AsInEx has grown to 70,000 unique organic substances. Samples are suppliable in 96-well plates and vials. NMR spectra upon request. Free database on CD disks. Contact: Oleg Darakov http://www.asinex.com From owner-repertoires@net.bio.net Wed Nov 05 22:00:00 1997 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.molecules.repertoires Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 6 Nov 1997 11:03:01 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 236 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <199710281000.CAA03383@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. From owner-repertoires@net.bio.net Wed Nov 05 22:00:00 1997 Path: biosci!biosci!not-for-mail From: rakonj@rockvax.rockefeller.edu (Jasna Rakonjac) Newsgroups: bionet.molecules.repertoires Subject: Anti c-myc tag antibody Date: 6 Nov 1997 11:03:39 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 23 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear all, I wonder whether anyone knows how to get or where to buy the anti c-myc tag mAb. Best regards and many thanks Jasna Rakonjac Zinder-Model Laboratory The Rockefeller University 1230 York Avenue, Box 24 New York, NY 10021 U.S.A. Phone: 212-327-8649 Fax: 212-327-7850 e-mail: rakonj@rockvax.rockefeller.edu From owner-repertoires@net.bio.net Sun Nov 09 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Antoine VEKRIS Newsgroups: bionet.molecules.repertoires Subject: Immunolocalisation of fd phage Date: 10 Nov 1997 00:15:55 -0800 Organization: Molecular pathology and gene therapy laboratory Lines: 57 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <3464765A.7A29@pmtg.u-bordeaux2.fr> Reply-To: Antoine.Vekris@pmtg.u-bordeaux2.fr NNTP-Posting-Host: net.bio.net > Immunolocalisation of fd phage > From: Emily Scott I am trying to screen various fd phage strains.... I really need a positive control of "just phage" but don't know how this would be done. Can I just dry a solution of phage onto a slide and stain this? Has anyone tried anything similar with any success? Hello Emily, and everybody out there, Positive controls allowing testing of the entry of the phage in permeabilised cells may be obtained as follows : a) Fix/permebilise cells with 4% PFA, 0.5% glutaraldehyde, 0.3 % triton X-100 in PBS, pH 7.2 for 1 h b) rince with PBS twice c) incubate with any phage suspention in PBS (! avoid carrier proteins they are lethal for the assay !) for 15 min d) saturate with 1 mg/ml BSA for 15 min e) ... proceed as usaually for the phage immunodetection The glutaraldehyde in the first step add free aldehydes on the sample. Phages are covalently trapped by them. Saturation with BSA is essential to avoid trapping the primary antibody. Confocal microscopy allows to estimate the diffusion of the phages into the cell. For antigens not aailable in the vicinity of the outer membrane further permeabilisation of the cells following fixation may be obtained by SDS 1 % in PBS, at RT, for 1-72 hours. An alternative using less optimisation steps is the inclusion of the cells pellet into parrafin and the preparation of sections of 5 micrometers. I have use this for simultaneous analysis of 24 samples on a single section. This allow comparisons of different cell lines/ a phage suspension interactions. a) suspend cells in 4 % PFA, 0.3 % triton X-100 in PBS, pH 7.2 for 1 h b) wash cells twice with PBS, collect by centrifugation at 400 g any resuspend by inversion of the tube rather then pippeting. c) following the second centrifugation carrefully eliminate the supernatant using a capillary d) resuspend the cells in melted parrafin (keep the tube HOT!), 10E+7 per 100 microliters. Pour the cell suspension in a preformed parrafin bed. e) allow to solidify and prepare sections. process as usually for immunohistrochemistry. Well, good luck and greetings from Bordeaux -- Antoine VEKRIS Molecular pathology and gene therapy laboratory University of Bordeaux II 146, rue Leo Saignat FR-33076 Bordeaux cedex Phone +33 (0)5 57571373, fax +33 (0)5 56983348, e-mail: vekris@u-bordeaux2.fr From owner-repertoires@net.bio.net Wed Nov 12 22:00:00 1997 Path: biosci!biosci!not-for-mail From: lpss@unixg.ubc.ca (Alex Chang) Newsgroups: bionet.molecules.repertoires Subject: phagemid vector Date: 13 Nov 1997 05:12:46 -0800 Organization: University of British Columbia, Vancouver, B.C., Canada Lines: 19 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <64doca$eet$1@nntp.ucs.ubc.ca> NNTP-Posting-Host: net.bio.net I'd like to obtain the phagemid vector pCANTAB6 (the one with a hexhistidine tag) or equivalent for expressing human ScFv library (Pharmacia's vector has a E-tag). It will be great if someone has that vector could send it to me. Thanks in advance. Alex Chang Pathology University of British Columbia 2222 Health Sciences Mall Vancouver, B.C. V6T 1Z3 Canada achang@hivnet.ubc.ca From owner-repertoires@net.bio.net Wed Nov 12 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Andrew Wallace Newsgroups: bionet.molecules.repertoires Subject: Re: Anti c-myc tag antibody Date: 13 Nov 1997 09:28:32 -0800 Organization: Queens University Belfast Lines: 34 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <346B0D1A.21A0@qub.ac.uk.see.signature> References: Reply-To: a.wallace@qub.ac.uk.see.signature NNTP-Posting-Host: net.bio.net Jasna Rakonjac wrote: > > Dear all, > I wonder whether anyone knows how to get or where to buy the anti > c-myc tag mAb. > > Best regards and many thanks > > Jasna Rakonjac > Zinder-Model Laboratory > The Rockefeller University > 1230 York Avenue, Box 24 > New York, NY 10021 > U.S.A. > Phone: 212-327-8649 > Fax: 212-327-7850 > e-mail: rakonj@rockvax.rockefeller.edu You can buy the 9E10 anti-myc tag monoclonal from Boehringer. There may be other sources also but we have found the Boehringer material to work well in our experiments. Andrew P.S. Usual disclaimer, no affiliation to Boehringer -- - note antispam feature in return address. My real address is: ================================================================== Andrew Wallace,Ph.D., Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html ================================================================== From owner-repertoires@net.bio.net Mon Nov 17 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Bill Town Newsgroups: bionet.molecules.repertoires Subject: ChemWeb's first virtual lecture Date: 18 Nov 1997 01:23:26 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 75 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <199711171846.SAA17106@cursci.co.uk> Reply-To: billt@CURSCI.CO.UK NNTP-Posting-Host: net.bio.net Register now for ChemWeb's first virtual lecture! We invite you to join us at the very first ChemWeb virtual lecture, "Great attractors in the chemical chaos of the Internet", which will be given by Dr. Henry Rzepa, from Imperial College, London, UK. The lecture will take place on Wednesday December 3rd, 1997, at 16:00 - 18:00 GMT in ChemWeb's Chemistry Auditorium [http://www.vei.co.uk/chemweb/]. Registration is required. The lecture is free to ChemWeb members. Henry Rzepa comments: "In my talk, I will introduce what I believe are some of the innovations over the last four years which the infra-structure of the Internet has spawned. These include electronic discussion lists, "active" conferences, new "third generation" electronic journals , impressive resources seen particularly in the area of bio-informatics, the fruits of the substantial labours in preparing new pedagogic materials and virtual courses, the new genre of "one-stop" chemical malls or clubs which attempt to integrate all these resources, and perhaps the experiment which we are all about to participate in of virtual talks. Important questions remain. How can we index and search for the seam of quality, with some 3000+ chemical web sites now operating? Is there yet an accepted mechanism for peer review and scholarly recognition in this medium? With an estimated half-life of a URL being around 6 months, are there not serious concerns about the permanence of materials? Copyright remains an almost intractible issue. Privacy is another thorny problem. The bottom line is can we do our science better with the help of the Internet or does it simply represent a source of global information pollution? Perhaps after a global brain-storming session, we can begin to see how the structure behind the chaos can be teased out, if we think it exists at all!" The format for the lecture is as follows: 16.00-16.30 Presenter - Henry Rzepa 16.30-17.00 Panel Member Presentations 17.00- 17.30 Presenter/Panel Discussion 17.30- 18.00 Open discussion (18.00- Discussion continues in Virtual Bar ...) Dr Wendy Warr will be moderating the lecture, and will be available to answer any questions you may have throughout the lecture. Panel members are: Oliver Kappe Peter Murray-Rust Mike Hann Steven Bachrach Steve Heller Chris Adams I look forward to meeting you at the lecture, Best regards Bill Town Chief Operating Officer ChemWeb, Inc. http://ChemWeb.com PS: don't forget to invite your colleagues to join us! PPS Apologies for multiple postings to discussion lists From owner-repertoires@net.bio.net Tue Nov 18 22:00:00 1997 Path: biosci!biosci!not-for-mail From: "Yi Zhang (BIO)" Newsgroups: bionet.molecules.repertoires Subject: Re: Anti-M13 phage antibody (mAB) needed. Date: 19 Nov 1997 02:21:39 -0800 Organization: University of South Florida Lines: 22 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <64sma3$hoq@mserv1.dl.ac.uk> Reply-To: yzhang1@chuma.cas.usf.edu NNTP-Posting-Host: net.bio.net Hi there, I remembered I saw it somewhere that some company out there sells Mab against gVIII antibody and gIII as well :-). It may be from New England Biolabs or pharmacia :-). I am not so sure now :-). Check NEB to see if they have it. Good luck ************************************************* * ___ Yi (Ian) ZHANG, * | | / IP2CMB, IBS * --- |/ USF, Tampa, FL * |__---- Web site: http://www.med.usf.edu/~yzhang/ * | |\ * | |/\ ************************************************* `` From owner-repertoires@net.bio.net Tue Nov 18 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Christoph Winter Newsgroups: bionet.molecules.repertoires Subject: Re: Anti-M13 phage antibody (mAB) needed. Date: 19 Nov 1997 02:24:54 -0800 Organization: MolGen Lines: 67 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <64smdb$hsu@mserv1.dl.ac.uk> Reply-To: chw@sirius.mgen.uni-heidelberg.de NNTP-Posting-Host: net.bio.net Hi! >Does anybody in this group know the any source of anti-M13 or fd phage >monoclonal antibodies? Any information lead to this will be greatly >appreciated. Try . ------------- Phone: +49 6221 8278 0 Fax: +49 6221 827823 email: info@www.progen.de mail: PROGEN Biotechnik GmbH Maa_strasse 30 D-69123 Heidelberg Germany ------------- Mouse monoclonal Filamentous fd, F1, M13 Phages Clone: B62-FE2 IgG2b purified IgG / supernatant The antibody binds to pVIII major coat protein of filamentous phages of the fd, F1 and M13 group. Suitable for the identification of recombinant phages in antigen and antibody-phage systems and phage ELISA. Reactive with: filamentous phages fd, F1, M13 Suitable for: ELISA, WB, immune electron microscopy Dilution: 1:500 for 107 phage particles (61097) Cat.No. Size 61097 100 5g purified (lyoph.) 65197 1 ml supernatant -------------------- Regards Christoph From owner-repertoires@net.bio.net Wed Nov 19 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Qiaoping Yuan Newsgroups: bionet.molecules.repertoires Subject: Anti-M13 phage antibody (mAB) needed. Date: 20 Nov 1997 13:32:35 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <64v93p$k2d@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Hi, there: Does anybody in this group know the any source of anti-M13 or fd phage monoclonal antibodies? Any information lead to this will be greatly appreciated. Qiaoping Yuan Michigan State University From owner-repertoires@net.bio.net Wed Nov 19 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Kajiwara Hideyuki Newsgroups: bionet.molecules.repertoires Subject: Re: Anti-M13 phage antibody (mAB) needed. Date: 20 Nov 1997 13:32:31 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <64v96s$k8j@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net It can purchase from Sigma. -- _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ Hideyuki Kajiwara National Institute of Agrobiological Resources Dept. of Biotechnology, Lab. of Gene Engineering E-mail: kajiwara@abr.affrc.go.jp Fax: 81-298-38-7073 _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ From owner-repertoires@net.bio.net Mon Nov 24 22:00:00 1997 Path: biosci!biosci!not-for-mail From: M.Foley@latrobe.edu.au (Dr Michael Foley) Newsgroups: bionet.molecules.repertoires Subject: random peptide libraries Date: 25 Nov 1997 02:03:24 -0800 Organization: La Trobe University Lines: 77 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <65d7or$pc7$1@news.latrobe.edu.au> NNTP-Posting-Host: net.bio.net A couple of weeks ago there was a discussion on which phage displayed random peptides are the best. I don't want to make any claims, but I just thought I'd let you know some of our experiences with two libraries that are in the public domain. Our lab has recently begun to screen phage displayed random peptide libraries with monoclonal antibodies to two malaria proteins. We have isolated several peptides which mimic the antigens used to elicit the original antibody. An antibody to one malarial antigen was used to pan two separate phage displayed peptide libraries (a 15mer from George Smith, Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211 USA; and a 17mer from Jamie Scott, Institute of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada V5A 1S6). The isolated clones were sequenced and were found to be unrelated peptide sequences although most of them shared a common amino acid motif within the different amino acid contexts The common motif contained a di-acidic repeat that resembles the parent antigen. Similar motifs were detected in peptides from the two libraries. Characterisation of these and other clones using ELISA, dot blotting demonstrates that the peptides bind specifically to the antibody used to isolate them and that recombinant antigen competes for binding. In addition rabbits immunised with the recombinant malaria antigen produce antibodies that recognise the peptides. We conclude that we have isolated mimotopes and are presently immunising mice with the peptides to confirm that the immune serum generated recognises the antigen in situ in the parasite. The George Smith library is five copies of the peptide fused to the gene3 protein and the Jamie Scott is fused to the gene8 protein so in this case there is several hundred copies. In addition the Jamie Scott library has a conserved cysteine as the second last amino acid. Both these libraries should allow the selection of different shapes even when panned with the same molecule. We find that they are extremely useful and having both these libraries increases the size of the pool of molecular shapes that we can search. I should also say that both George and Jamie are very helpful supply useful protocols. We have panned these libraries on a couple of other antibodies and from what we have looked at so far we get a similar sort of thing ie different peptides but a common motif within the amino acid sequence. Hope this has been of some use to somebody. By the Way does anyone know of any papers or occasions when the structure of a short peptide ~ 15amino acids has been worked out by NMR or some other technique? From owner-repertoires@net.bio.net Thu Nov 27 22:00:00 1997 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.molecules.repertoires Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 28 Nov 1997 02:17:11 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 236 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <199711281000.CAA20999@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. 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From owner-repertoires@net.bio.net Thu Nov 27 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Andrew Wallace Newsgroups: bionet.molecules.repertoires Subject: Re: random peptide libraries Date: 28 Nov 1997 02:13:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 133 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Michael, Thank you for your valuable observations and comments. I agree with you that it is a good idea to screen with a variety of libraries to maximise the possibility of finding interesting hits within the conformational space accessed by each library. No two phage libraries will give exactly the same result, but similar peptides are probably displayed in each different system such that a consensus about the "best binder" can be obtained. I think that the chances of finding a small (<15mer) peptide which adopts a particular structure in solution are quite low and I do not know of any published examples (apart from anecdotal rumours). On the other hand, I am sure there are several examples of small peptides adopting a particular structure upon binding to their cognate ligate. e.g., this is what I found concerning the Scripps antibody which recognises a linear epitope from influenza hemagglutinin TI: STRUCTURAL EVIDENCE FOR INDUCED FIT AS A MECHANISM FOR ANTIBODY- ANTIGEN RECOGNITION AU: RINI_JM, SCHULZEGAHMEN_U, WILSON_IA JN: SCIENCE, 1992, Vol.255, No.5047, pp.959-965 TI: CRYSTAL-STRUCTURE OF A PEPTIDE COMPLEX OF ANTIINFLUENZA PEPTIDE ANTIBODY-FAB-26/9 - COMPARISON OF 2 DIFFERENT ANTIBODIES BOUND TO THE SAME PEPTIDE ANTIGEN AU: CHURCHILL_MEA, STURA_EA, PINILLA_C, APPEL_JR, HOUGHTEN_RA, KONO_DH, BALDERAS_RS, FIESER_GG, SCHULZEGAHMEN_U, WILSON_IA JN: JOURNAL OF MOLECULAR BIOLOGY, 1994, Vol.241, No.4, pp.534-556 * TI: STRUCTURE OF ANTIPEPTIDE ANTIBODY COMPLEXES AU: WILSON_IA, GHIARA_JB, STANFIELD_RL JN: RESEARCH IN IMMUNOLOGY, 1994, Vol.145, No.1, pp.73-78 * TI: DETAILED ANALYSIS OF THE FREE AND BOUND CONFORMATIONS OF AN ANTIBODY - X-RAY STRUCTURES OF FAB 17/9 AND 3 DIFFERENT FAB-PEPTIDE COMPLEXES AU: SCHULZEGAHMEN_U, RINI_JM, WILSON_IA JN: JOURNAL OF MOLECULAR BIOLOGY, 1993, Vol.234, No.4, pp.1098-1118 Best regards, Andrew On 25 Nov 1997 02:03:24 -0800 Dr Michael Foley wrote: > A couple of weeks ago there was a discussion on which phage displayed random > > peptides are the best. I don't want to make any claims, but I just thought I'd > > let you know some of our experiences with two libraries that are in the public > > domain. > Our lab has recently begun to screen phage displayed random peptide libraries > > with monoclonal antibodies to two malaria proteins. We have isolated several > > peptides which mimic the antigens used to elicit the original antibody. An > > antibody to one malarial antigen was used to pan two separate phage displayed > > peptide libraries (a 15mer from George Smith, Division of Biological Sciences, > > University of Missouri, Columbia, Missouri 65211 USA; and a 17mer from Jamie > > Scott, Institute of Molecular Biology and Biochemistry, Simon Fraser > > University, Burnaby, BC, Canada V5A 1S6). The isolated clones were sequenced > > and were found to be unrelated peptide sequences although most of them shared a > > common amino acid motif within the different amino acid contexts The common > > motif contained a di-acidic repeat that resembles the parent antigen. Similar > > motifs were detected in peptides from the two libraries. Characterisation of > > these and other clones using ELISA, dot blotting demonstrates that the peptides > > bind specifically to the antibody used to isolate them and that recombinant > > antigen competes for binding. In addition rabbits immunised with the > > recombinant malaria antigen produce antibodies that recognise the peptides. We > > conclude that we have isolated mimotopes and are presently immunising mice with > > the peptides to confirm that the immune serum generated recognises the antigen > > in situ in the parasite. > The George Smith library is five copies of the peptide fused to the gene3 > > protein and the Jamie Scott is fused to the gene8 protein so in this case there > > is several hundred copies. In addition the Jamie Scott library has a conserved > > cysteine as the second last amino acid. Both these libraries should allow the > > selection of different shapes even when panned with the same molecule. We find > > that they are extremely useful and having both these libraries increases the > > size of the pool of molecular shapes that we can search. I should also say that > > both George and Jamie are very helpful supply useful protocols. We have panned > > these libraries on a couple of other antibodies and from what we have looked at > > so far we get a similar sort of thing ie different peptides but a common motif > > within the amino acid sequence. > > Hope this has been of some use to somebody. > > By the Way does anyone know of any papers or occasions when the structure of a > > short peptide ~ 15amino acids has been worked out by NMR or some other > > technique? > > > ------------------------------------------------------------------ Andrew Wallace, Ph.D, Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html From owner-repertoires@net.bio.net Sun Nov 30 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Alicia Gomez Roig Newsgroups: bionet.molecules.repertoires Subject: pHen and purifications Date: 1 Dec 1997 11:14:21 -0800 Organization: MERCK-LBI (Barcelona) Lines: 15 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I would like to ask the group something: I would like to know who is working with pHEN vector for diferencial display , and also how you purified the clones that you find interesting for your research. I am working with the Pharmacia kit (HiTrap Anti-E tag) but the recovery is very low. Thank you very much for your time. Sincerely yours Alicia.Gomez@merck.de From owner-repertoires@net.bio.net Sun Nov 30 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Andrew Wallace Newsgroups: bionet.molecules.repertoires Subject: Meetings and Courses 1998 Date: 1 Dec 1997 02:35:40 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 58 Sender: daemon@net.bio.net Approved: A.Wallace@Queens-Belfast.AC.UK Distribution: world Message-ID: <65u367$fvi@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Meetings and courses on combinatorial libraries and phage display. ***************************************************************** _________________________________________________________________ *** MEETINGS *** _________________________________________________________________ __________________________________________________________________ The 8th Cyprus Conference on New Methods in Drug Research Limassol, Cyprus May 4-9, 1998 Conference Executive: Chair: Alex Makriyannis (U. Conn. Storrs, USA) Committee: N. Castagnoli (USA), J.P. Devlin (USA), U. Hacksell (Sweden), L.H. Hurley (USA) and M. Abou-Gharbia (USA) Conference Location Time and Registration: The Conference will take place from Monday, May 4th through Friday, May 9th 1998 at the five-star Hawaii Beach Hotel, Limassol, Cyprus. Attendance is restricted to 150 participants to encourage interaction. Registration fees ($595 per person; $395 for academics) includes attendance at all sessions and morning and afternoon breaks, but not meals. Sponsor: The European Federation of Medicinal Chemistry Contact: ARRT International, Inc., P.O. Box 1838, New Milford, CT 06776, USA Telephone: 860-355-5195; Facsimile: 860-355-5975; e-Mail: arrtintl@prodigy.net _________________________________________________________________ *** COURSES *** _________________________________________________________________ Phage Display of Combinatorial Antibody Libraries November 11 - 24 Organised by: C. Barbas, D. Siegel, G. Silverman Contact: Cold Spring Harbor Laboratory The Meetings & Courses Office PO Box 100, 1 Bungtown Road Cold Spring Harbor, NY 11724-2213 Phone: 516-367-8346 Fax: 516-367-8845 Email: meetings@cshl.org _________________________________________________________________ ------------------------------------------------------------------ Andrew Wallace, Ph.D, Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html