stable protein complex during SDS-PAGE

Dr E. Buxbaum EB15 at le.ac.uk
Mon Mar 2 13:09:01 EST 1998


brendon wrote:
> 
> Thomas <urbig at biokemi.su.se> wrote in article
> HI - I HAVE HAD THE SAME TROUBLE IN THE PAST. YOU CAN DENATURE YOUR
> PROTEINS IN 6M GUANIDINE HCL, 5 mM EDTA, pH 8.1. FROM THERE, REDUCE WITH
> DITHIOTHREITOL AND CARBOXMETHYLATE.  GOOD LUCK

Guanidinum HCl may not be the ideal reagent for use in electrophoresis.
Urea, being non-charged, might be the better choice. The other little
trick I found usefull is to incubate the sample in Laemli buffer for 30
min at 37 degrees, instead of boiling them.



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