From owner-drosophila@net.bio.net Fri Nov 10 22:00:00 1995 Path: biosci!PSU.EDU!gxt5 From: gxt5@PSU.EDU (Graham Thomas) Newsgroups: bionet.drosophila Subject: Re: help with antibody Date: 11 Nov 1995 10:19:36 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 42 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Artur Kania wrote >.....Is there a way to modify the fixation to include a detergent or something >which will denature my antigen ? Should I use this detergent throughout >the Ab staining protocol ? Has anyone encontered this problem before (and >solved it) ? >.... I believe that there is a mixed-micelles technique that I think may help in this kind of situation. Add SDS to your detergent at 1/4-1/5th the concentration of your triton - e.g. 0.02-0.033% SDS + 0.1% Triton. Unfortunately I have no citation to give you; however, my notes indicate that this method came originally (or at least to the person who told me) from Larry Gerace at the Scripps so you might want to contact him if you want to try this for more details (- and convey my sincere apologies if this attribution is incorrect). I solved my problems in other ways and so I cannot offer you the benefit of extensive experience in it's use. Maybe this will jog someones memory out there. Good Luck Graham FliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFlies l i We have a new graduate school opportunity. Please TRY e THIS URL and PASS IT ON to any aspiring graduate students. s http://www.bmb.psu.edu/thomas/cdb/cdbhome.htm F l i Graham Thomas, e Departments of Biology and Biochemisty/Molecular Biology, s The Pennsylvania State University, F 208 Mueller Laboratory, l University Park, PA, 16802 i e Email GXT5@ PSU.EDU s Telephone (814)-863-0716 F Fax (814)-865-9131 l WWW http://millreef.mcb.psu.edu/thomas i esFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFliesFli .