Alternatives to Laemli?
Michael L. Sullivan
mlsulliv at facstaff.wisc.edu
Tue Feb 17 11:47:55 EST 2004
>As for the original post, I don't think SDS-based buffer would work for
>studying protein/RNA complexes at all, simply because SDS would denature the
>protein. There are non-denaturing buffer systems, like Tris-borate-EDTA
>(TBE) that work fine for that purpose.
I think another approach that people use when studying RNA binding
proteins is to do UV crosslinking/label transfer type protocols.
With that approach, a radio labeled RNA target is incubated with a
protein extract, RNA is UV crosslinked to the protein, and then
excess RNA is degraded enzymatically. This leaves proteins with
(hopefully) small pieces of labeled RNA attached which can then be
RUN on a regular SDS-PAGE gel and detected by autoradiography..
Michael L. Sullivan, Ph.D
Research Plant Molecular
U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706
(608) 264-5397 Phone
(608) 264-5147 Fax
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