enzymatic digests on PVDF bound samples-coomassie problem

Suzanne Perry-Riehm (perry@interchange.ubc.ca)
Wed, 1 Dec 1999 13:12:46 -0800 (PST)

Does anyone out there who performs enzymatic digests on PVDF bound samples
have any solutions to the coomassie stain which accompanies the samples?

I regularly receive samples in which the PVDF bound
samples are grossly over-stained with coomassie blue. After performing the
digest and taking the sample through the speed vac I get about
20 micro-liters of
dark blue sludge. This is hell on the PE ABI 173A microblotter and
associated cHPLC, not to mention the chromatograms.

Is there a de-stain step I could use that wouldn't affect the protein
prior to the digestion? Is there a pre-column that might also be useful
for removing the stain prior to the cHPLC column?

Any thoughts on this are greatly appreciated!

Best Regards,
Suzanne Perry-Riehm

Suzanne C. Perry, Research Associate
NAPS Protein Sequencing and Peptide Mapping Laboratory
Biotechnology Department, University of British Columbia
CANADA
phone:(604)822-9688( direct) or (604)822-9662 (main) Fax:(604)822-0676
email:perry@interchange.ubc.ca
Visit our web page at : www.biotech.ubc.ca/naps/napshome.htm