reprobe western blots

kramerse at wmvx.lvs.dupont.com kramerse at wmvx.lvs.dupont.com
Fri Jan 6 11:55:40 EST 1995


In article1995Jan6.025507.:
>In <anon-0501951543180001
                       
>>In article <3eg2t9$8hl at neuro.usc.edu>, william at neuro.usc.edu (Fiberman) wrote:
>
>>> Hello, Netters,
>>> 
>>> Can a western blot filter be "reprobed" like a Southern blot filter?  If
>>> so, what is the best way to strip the original antibody?  I am using a
>>> secondary antibody conjugated to alkaline phosphatase, and visualizing
>>> with addition of color substrates NBT and BCIP.  I would appreciate your
>>> response.
>>> 
>>> -fm
>
>
Stripping of antibodies from membranes has been described for 125-I
labeled proteins and the extension of theis technique to chemiluminescence
substrates is feasible.

Reference:
Kaufman,et al., Analytical Biochem., 161:89-95,(1987)

Removing chromogenic substrates is difficult, I don't reccommend it.


>                                 
>--                                         
>Tom Thatcher                          | You c
>University of Rochester Cancer Center | and h



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