eluting proteins from gels

Vladimir Rotrekl rotrekl at chemi.muni.cz
Mon Jun 1 03:05:34 EST 1998


Karen spink wrote:

> Recently, i have been dabbling in a bit of protein purification and on
> a
> coomasie gel i have only a few proteins, one in particular is stronger
> than
> the others. I wish to show that this stronger band is the protein
> responsible for activity I am detecting in the fraction. Is it
> feasible to
> simply elute this band from the SDS gel and assay this for activity,
> i.e.
> is there a simple way of eluting the protein directly from the gel
> with a
> good chance of renaturing the protein?
>
> Thanks in anticipation
>
> Karen Spink
>
> E. mail pdxkgs at pdn1.gene.nottingham.ac.uk

   I havent heard any method where You can get activity after SDS.
Better to try native polyacrylamide gel. Then just cut out the band of
interest and check the activity without getting rid of polyacrylamide.
But!!! knowing nothing about Your proteine  it is hard to predict Your
chances. For some enzyme activities there are available activity gel
staining procedures (e.g. for gycosidases).

good luck Vladimir Rotrekl
               Dept of biochemistry
               Masaryk University, Kotlarska 2, 61137 Brno, CZECH REP.
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