protein/peptide elution from nitrocellulose/PVDF

Dennis Takayesu takayesu at medcor.mcgill.ca
Thu Jun 20 07:58:55 EST 1996


I've been transferring proteins to nitrocellulose and PVDF from SDS-Page
gels, excising the protein bands and digesting the proteins with
trypsin,cyanogen bromide, etc. I want to make certain that I'm getting
most of the resulting peptides off of the membrane. The protein has
palmitate and myristate attached co- and post-translationally, so some of
the resulting peptides from the digestion may be hard to remove from the
membrane.
The resulting eluates are analyzed by reverse-phase HPLC. 

Does anyone have any protocols that address this situation i.e. removal of
proteins or peptides form nitrocellulose or PVDF membranes (especially
fatty acid modified species)?

Thanks
Dennis

-- 
Richard C. Marcellus
Dept. of Biochemistry
McGill University
Montreal, Canada
marcellus at medcor.mcgill.ca



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