SDS-PAGE

Nikolai Chitaev nikolaic at VISAR.WUSTL.EDU
Mon Nov 27 17:16:02 EST 1995


> From: lpss at unixg.ubc.ca (Alex Chang)
> Subject: SDS-PAGE
> Date: 27 Nov 1995 19:50:26 GMT
> Hi, everyone:
> 
> Something wired happened to my SDS-PAGE gel. I did SDS-PAGE many times
> before, but haven't done it for a while. Recently, when I adapted my old
> protocal for a mini-gel, it takes hours for the separating gel to
> solidify and even worse, the stacking gel solidified poorly and the wells
> did not contain the samples very well as the result of it.
>... <<much deleted>>...
> Alex Chang
> Pathology
> University of British Columbia

Poor (slow) polimerization of acrylamide means that your APS is dead. If the APS is
freshly made and problem persists - dead TEMED (instead to adding more TEMED buy new).
After the moleculer net is formed the resulting gel squeezes excess of water, since the 
gel now ocupies less volume then two of the components together. The amount of water 
relevant to the T, i.e. proportion of Bis-acrylamide/Acrylamide.
Another factors which affect the rate of polymerization could be the purity of reagents 
used, as a potential inhibitors of APS as well as purity of the glasware.
cheers
Nikolai
http://128.252.119.253






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