From baorui_ren from hotmail.com Mon Aug 4 21:30:59 2008 From: baorui_ren from hotmail.com (=?gb2312?B?sPzI8Q==?=) Date: Tue Aug 5 11:14:14 2008 Subject: [Protein-analysis] HELP! protein-protein cross-linking Message-ID: From: baorui_ren@hotmail.comTo: methods@net.bio.netSubject: HELP! protein-protein cross-linkingDate: Tue, 5 Aug 2008 02:28:20 +0000 Hello, AllWe have a problem in the chemical cross-linking experiment for protein-protein interaction. We have solved a protein complex structure, and we want to confirm their interaction by cross-linking. But i am a new comer in this field, the initial test was failed. Protein A is 35kd, Protein B is 14kd, but after cross-linking the band representing 49kd is very very weak in our SDS-PAGE comparing with the inact Protein A and Protein B in thesame lane. The condition we used is : add DSS to protein sample containing protein A and protein B and incubated for 1 hour under 4 degree, and then terminated it with 1M tris for 30 min. The samples were loaded on SDS-PAGE. We have tried different concentrations of DSS (0.1mM-10mM) and protein sample (20uM-1mM). PS: from the complex structure , we have found several Arg residues in both protein were located in the interface, so we use the DSS for initial test.Protein A trends to form dimer, but we could not even got the dimer bands after cross-linking. I would like to hear any suggestion from you.Thanks! From sumida from bbri.org Mon Aug 11 18:27:16 2008 From: sumida from bbri.org (jps) Date: Mon Aug 11 19:30:45 2008 Subject: [Protein-analysis] Re: HELP! protein-protein cross-linking References: Message-ID: <4ec89863-a42e-413b-88b8-92b728f05991@t54g2000hsg.googlegroups.com> I suppose some intial questions would have to do with some basic issues which I'm sure you have been careful about but one needs to ask them anyway in order to move forward. 1) Is your buffer free of reactive amines ie. presumably it is not a tris or imidazole type buffer? 2) Is your reagent fresh? 3) Are you working in a pH range 7-9? And are you sure? Second order concerns would have to do with the site accessibility of DSS to the reactive amines on the protein. Do you know if this site could be sterically blocked? How does trypsin interact with your protein? Chances are if trypsin does cleave at the reactive site or if the rate of cleavage at the reactive site is slowed, there are steric issues with the accessibity of the probe to the reactive site. Do you know if there are charged groups that could result in a coulombic interaction with your probe? Of course these latter more interesting queastions should only be addressed after the less interesting but more important questions are satisfactorily answered. =B0=FC=C8=F1 wrote: > From: baorui_ren@hotmail.comTo: methods@net.bio.netSubject: HELP! protein= -protein cross-linkingDate: Tue, 5 Aug 2008 02:28:20 +0000 > > > Hello, AllWe have a problem in the chemical cross-linking experiment for = protein-protein interaction. We have solved a protein complex structure, an= d we want to confirm their interaction by cross-linking. But i am a new com= er in this field, the initial test was failed. Protein A is 35kd, Protein B= is 14kd, but after cross-linking the band representing 49kd is very very w= eak in our SDS-PAGE comparing with the inact Protein A and Protein B in the= same lane. The condition we used is : add DSS to protein sample containing = protein A and protein B and incubated for 1 hour under 4 degree, and then t= erminated it with 1M tris for 30 min. The samples were loaded on SDS-PAGE. = We have tried different concentrations of DSS (0.1mM-10mM) and protein samp= le (20uM-1mM). PS: from the complex structure , we have found several Arg r= esidues in both protein were located in the interface, so we use the DSS fo= r initial test.Protein A trends to form dimer, but we could not even got th= e dimer bands after cross-linking. I would like to hear any suggestion from= you.Thanks! From dogoyi from mail.uonbi.ac.ke Sun Aug 17 06:27:32 2008 From: dogoyi from mail.uonbi.ac.ke (dogoyi@mail.uonbi.ac.ke) Date: Sun Aug 17 16:40:09 2008 Subject: [Protein-analysis] Fungal DNA isolation Message-ID: <5accb7498ee162ac677539424bb8079d.squirrel@mail.uonbi.ac.ke> Hi, Having real difficulties getting DNA from some fungal isolates. We have tried a number of techniques without much success. Anybody out there who has a non-kit technique that works well? Dr. D. O. Ogoyi Department of Biochemistry University of Nairobi P.O. BOX 30197, 00100 NAIROBI, KENYA ----------------------------------------- UNIVERSITY OF NAIROBI IS NOW ISO CERTIFIED The University of Nairobi is committed to providing quality services to all its clients. The University will monitor and review its quality performance from time to time through an effective implementation of the Quality Management System based on ISO 9001:2000 standard. University of Nairobi Website: http://www.uonbi.ac.ke/ -------------------------------------------- From sticher from bioc.unizh.ch Mon Aug 18 10:36:32 2008 From: sticher from bioc.unizh.ch (Patrick Sticher) Date: Mon Aug 18 16:04:47 2008 Subject: [Protein-analysis] 6th NCCR Symposium on New Trends in Structural Biology: New Program / Registration deadline August 29 Message-ID: <48A99700.6000404@bioc.unizh.ch> Dear colleagues, 6th International NCCR Symposium on New Trends in Structural Biology 8 + 9 September 2008, University of Z?rich, Z?rich, Switzerland NEW PROGRAM. The opening lecture will now be given by Gebhard Schertler about "Structure of the stress hormone receptor: beta1 adrenergic receptor". Confirmed plenary lecturers are Markus Gr?tter, Stephen C. Kowalczykowski, Kaspar Locher, Keiichi Namba, Poul Nissen, Andrej Sali, Gebhard F.X. Schertler, Jeffrey Skolnick, and A. Joshua Wand. The meeting will be held together with the Annual Meeting of the Swiss Society for Crystallography. Speakers of the society are Clemens Schulze-Briese and Colin Nave. Please see the symposium website for the complete program. REGISTRATION SLOT EXPIRES AUGUST 29. Please note that online registrations can be accepted for only a few more days until August 29. INFORMATION AND REGISTRATION. www.structuralbiology.uzh.ch/symposium2008.asp Please do not hesitate to contact me anytime if you need further information (sticher@bioc.uzh.ch). With best regards, Patrick Sticher The NCCR Structural Biology is a research initiative of the Swiss Science Foundation. Its research encompasses the fields of recombinant protein technologies, macromolecular structure determination and computational biomolecular sciences with a special focus on membrane proteins and supramolecular assemblies/interactions. 19 research groups from Swiss Universities and Research Institutions participate in this network. www.structuralbiology.uzh.ch/ _________________________________ Dr. Patrick Sticher Moser NCCR Scientific Officer Institute of Biochemistry University of Z?rich Winterthurerstrasse 190 CH - 8057 Z?rich Phone +41 / (0)44 / 635 54 84 Fax +41 / (0)44 / 635 59 08 From dogoyi from mail.uonbi.ac.ke Sat Aug 16 11:02:20 2008 From: dogoyi from mail.uonbi.ac.ke (dogoyi@mail.uonbi.ac.ke) Date: Mon Aug 18 16:05:14 2008 Subject: [Protein-analysis] (no subject) Message-ID: <83da4aaad74b07e5f55a824bb5a3015b.squirrel@mail.uonbi.ac.ke> Hi, Having real difficulties getting DNA from some fungal isolates. We have tried a number of techniques without much success. Anybody out there who has a non-kit technique that works well? Dr. D. O. Ogoyi Department of Biochemistry University of Nairobi P.O. BOX 30197, 00100 NAIROBI, KENYA ----------------------------------------- UNIVERSITY OF NAIROBI IS NOW ISO CERTIFIED The University of Nairobi is committed to providing quality services to all its clients. The University will monitor and review its quality performance from time to time through an effective implementation of the Quality Management System based on ISO 9001:2000 standard. University of Nairobi Website: http://www.uonbi.ac.ke/ -------------------------------------------- From engelbert_buxbaum from hotmail.com Tue Aug 19 09:02:43 2008 From: engelbert_buxbaum from hotmail.com (Dr Engelbert Buxbaum) Date: Tue Aug 19 13:57:20 2008 Subject: [Protein-analysis] Re: request for reprint..it is very important References: <833c3f65-2e84-4e6c-8e5f-483d5a6e408a@k30g2000hse.googlegroups.com> Message-ID: Am 16.08.2008, 17:26 Uhr, schrieb shrinivas Dengeti : > if anybody could get this paper please mail to my email ID :- > vasu.nanobiotech91@gmail.com That would be a breach of copyright. The legal way to do this is to email the corresponding author and ask for a (electronic) reprint. From labmend from hotmail.com Sun Aug 31 13:57:29 2008 From: labmend from hotmail.com (Peter Billington) Date: Mon Sep 1 14:48:49 2008 Subject: [Protein-analysis] FW: FPLC Support Message-ID: Dear Sirs=2C Scientific Instrument Repairs Ltd=2C based in the UK=2C would = like to inform you that we offer an experienced maintenance and repair serv= ice for the full range of Amersham/Pharmacia FPLCTM instruments=2C many of = which are no longer supported by the OEM manufacturer. For a quotation ple= ase do not hesitate to call or e-mail. Contact: Peter Billington. tel: 0796= 0 445672 e-mail: labmend@hotmail.com =20 _________________________________________________________________ Win New York holidays with Kellogg=92s & Live Search http://clk.atdmt.com/UKM/go/107571440/direct/01/=