Protein and agarose gels

Christopher Blencowe cbb at convex.phazc.uni-heidelberg.de
Mon Dec 12 06:57:52 EST 1994


James J. Campanella (jjc4 at po.CWRU.Edu) wrote:
: Has anybody had any experience running protein on agarose
: gels? We know that you get very low resolution, but we're
: looking at high molecular weight (~150 KD) bands that are
: in very high abudance and we want to run a fast gel to see
: if they're present. What concentrationn of agarose is
: generally used for such a gel? Thanks.
: -- 
: "Sometimes the less travelled is less travelled for a reason...."
:                                        -Jerry Seinfeld

Concentration is not critical, because separation is done
mostly on the absolute charge/molecule, no size sieving as
done in PAGE. 

So optimisation should be done using different buffers (pH), 
gel concentration should be 1% as a rule of thumb for big
sized molecules.

Better make a gradient or non-gradient SDS-PAGE of 10% or less
%T (total acrylamide concentration).

Sincerly



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