Stripping Western blots
(by as9n from virginia.edu)
Fri Oct 26 12:31:59 EST 2007
You're right - better to describe the already tested methods. The
methods we have tried are:
1. 1% SDS in PBS overnight at 37degreesC. This has variable results
depending on the antibody, but doesn't result in much loss of protein.
2. 2% SDS, 100mM beta-mercaptoethanol, 62.5mM Tris pH 7.4, for 30
minutes either at 50 degreesC or at room temperature. This works for
blots of cultured cell lysates, but results in high loss of protein
so it is too harsh for blots of embryo protein.
We're looking at the degree of phophorylation of several proteins, so
we need to have good removal of antibody, but we have very little
protein, so we need to minimize protein loss.
More information about the Methods