Ponceau S staining of PVDF membranes?

Bill Nowatzke wnowatzk at ucs.indiana.edu
Mon Jan 31 18:57:53 EST 1994


I have been trying to blot a tryptic fragment from an 8% SDS PAGE to
BioRad PVDF membrane. I thought that my blot was unsuccessful after
attempting to visualize by Ponceau S. I could not even detect my MW
standards. I decided to coomassie stain after destaining and saw all of
the protein that I had thought I loaded. Any clues as to why my ponceau S
did not work? I diluted a fresh Sigam concentrate (20 mL) into 180 mL of
deionized water. I stained for 5 min and watch as the membrane destained
in 1% AcOH. At no time did I see anything resembling a protein band. This
problem has occured for the past two consecutive times that I have tried
the technique. Your comments are appreciated.



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