nondenaturating PAGE

Ulf Sommer usommer at biochem.uni-kiel.de
Thu Mar 19 12:11:12 EST 1998


Silke Beismann wrote:
> 
> Who can give me some hints?
> I want to carry out a nondenaturating PAGE. Neither me nor anyone else
> in my lab has ever done this before.
> What is the best buffer system? Can I cast the gel in the same slab gel
> casting device used for SDS-gels?
> What picture would one expect to find if I load the gel with two
> proteins I expect to form a complex? One is 23 kDa with a pI of 6.79 and
> a net charge of -2, the other one is 28 kDa with a pI of 5.06 and a net
> charge of -8.
> I will be happy about any help I can get!
> Silke

Hi Silke,
try your normal PAGE conditions without adding SDS in any buffer you
use. Maybe you have to vary the concentration of the resolving gel.
Normally, the complex should be slower, but find it out yourself.
And perhaps you may find something in a protein purification book in
your library?
I hope that helps.
	Ulf



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