PVDF blotting problem

Rob mbzrl at mbn1.biochem.nottingham.ac.uk
Thu Jul 16 04:53:56 EST 1998


Hi
I'm trying to blot a protein to PVDF for sequencing but i'm having
problems. The protein blots fine to nitrocellulose in Tris/glycine/methanol
at pH8.3, but will not leave the gel in CAPS or CAPS/SDS pH11. Has anyone
got experience of blotting what I assume to be a basic protein eg in 0.1%
acetic and blotting towards the cathode? Also, could I use the CAPS at a
lower pH eg. 9 or 10 (pKa 10.4)?
Thanks in advance
Rob Layfield
University of Nottingham



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