SDS-PAGE gel resolution

Oswaldo oswaldo at iats.csic.es
Thu Aug 3 05:41:43 EST 2000



Martin Thompson wrote:
> 
> I was wondering if anyone knows what kind of factors influence how
> 'fuzzy' bands are on a SDS-PAGE gel.  Currently I am having problems
> with this and would like to know how to increase the sharpness or
> resolution of these bands.

Assuming that you are using right procedures and reagents for casting
and running the gels, and that the reduction/denaturation of the samples
is correct, I'd suggest to try concentrating the samples so you load the
smallest volume possible. This usually produces much sharper bands.
Amicon's Microcon (no affiliation) concentrating membranes with a 10.000
Da cutoff are useful for this if your protein is heavier.

Other thing to try would be gradient gels. Hope that it helps,

Oswaldo






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