HELP: Stripping western blots

Manuel Simon ma at ABC.univie.ac.at
Thu Apr 28 10:12:09 EST 1994


In article <Coxopn.5wM at ucdavis.edu> ez005528 at rocky.ucdavis.edu (Kevin Morano) writes:
>From: ez005528 at rocky.ucdavis.edu (Kevin Morano)
>Subject: HELP: Stripping western blots
>Date: Wed, 27 Apr 1994 19:56:10 GMT

>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

High, 
I do not have an explanation for that but to my experience I can not strip Ab 
from certain membranes (Amersham). E.g. since I used routinely nylon material 
my yields of stripped Ab is reduced to 50% (or even less) of that what I used 
to get in fromer experiments with the same Ab (e.g. from Biorad´s 
NC-membranes).The second thing is, that I know Ab´s you cannot strip except 
using urea or heavy stuff like this. 

Manuel Simon



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