HELP: Stripping western blots

littgj at wmvx.lvs.dupont.com littgj at wmvx.lvs.dupont.com
Tue May 3 07:04:50 EST 1994


In article <Coxopn.5wM at ucdavis.edu>, ez005528 at rocky.ucdavis.edu (Kevin Morano) writes:
>I'm trying to do sequential probings of a western blot with different 
>antibodies and am having a tough time stripping the previous antibody 
>off. I have been using a protocol which is essentially an acid (pH 2.2) 
>glycine/NaCl/TX-100 wash, and it looks like less than 10% of the old 
>primary Ab is coming off. What do people use out there? By the way, I 
>develop with DUPont's Renaissance chemi. kit, so I could theoretically do 
>many probings.
>
>Thanks in advance!
>Kevin
>
>--
>---------------------------------------------------------------------------
>Kevin A. Morano
>Internet:kamorano at ucdavis.edu    
>Bitnet:kamorano at ucdavis   
>Section of Microbiology, University of California, Davis
Kevin,
I tried to get through to your kamorano at ucdavis.edu address
yesterday but got bounced! Annyway, we prepared a note for our
Biotech Update in the summer of 1993 which described the use of
the following to strip membranes: 100 mM 2-mercaptoehanol , 2% SDS,
62.5 mM Tris-HCl (pH 6.8) for 30 min. at 50 degrees C. 
If you want a copy of the note, send me your FAX #.
Jerry Litt (LITTGJ at WMVX.LVS.DUPONT.COM)



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