western blot stripping protocol

Anthony Martinez szsnoopy at chip.ucdavis.edu
Wed Jan 31 17:55:52 EST 1996


I am having problems stripping off my primary antibody off of my 
western blots.  I am using nitrocellulose and following NEN's stripping 
protocol that calls for a 30 minute incubation in 62.5 mM Tris-HCL pH 
6.8, 2% SDS, and 100 mM 2-mercaptoethanol at 50 degrees centigrade.  My 
results show that only the secondary antibody is being washed off and my 
primary is still there.  Any suggestions, comments or is there a better 
protocol to remove the primary antibody off nitrocellulose?

Thanks in advance.

Tony Martinez



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