Electrotransfer proteins for Edman sequencing

Emir Khatipov khatipovNO at NOuchicago.edu
Fri Nov 2 20:21:31 EST 2001

Hello All,
Could someone recommend me a good protocol for SDS-PAGE and subsequent
transfer of proteins onto PVDF membrane for subsequent Edman sequencing?

Which PVDF membrane did you use, how did you deal with non-polymerized
acrylamide that can modify proteins and impair sequencing, what transfer
buffers worked for you best, etc. Should I use PAGE buffers different from
Tris-glycine (Borate?), and do I have to care about glycine on Page stage at
all? I know that CAPS buffer is a good alternative for Tris glycine transfer
buffer, but I am concerned that high pH of the buffer (pH 11) will cause
modification of side chains of amino acids. In other words, I good detailed
working protocol would be very much appreciated, or pointers to published

Thank you.

University of Chicago
Radiation Oncology

More information about the Proteins mailing list

Send comments to us at biosci-help [At] net.bio.net