protein extraction from gel
Dr Engelbert Buxbaum
(by engelbert_buxbaum from hotmail.com)
Mon May 26 07:35:55 EST 2008
Am 22.05.2008, 11:25 Uhr, schrieb crivera <crivera from cibnor.mx>:
> to purifiy an enzyme, i try different methodologies (Chromatography:HIC,
> GF,IC), now im looking for a protocol to extract proteins from gel, I
> saw the message on internet about elute protein from gel, could you
> >please send me the refrence or protocol to do it.
This method has only limited applicability for enzyme isolation, as most
electrophoretic methods - in particular SDS PAGE - work with denatured
proteins. You could however try methods designed to keep the protein
happy, e.g. Blue native or CTAB PAGE, the latter of course without
SH-reagent in the sample buffer.
In principle, you cut the band out of the gel (blue native PAGE is nice as
you do not need to stain), homogenize the slice (passage through 27G
needle or Dounce homogenizer) and place the bits onto a piece of Muller
gauze into a tube with buffer. The end of the tube is closed with dialysis
membrane. Electrical current drives the protein toward this membrane.
Several manufactureres, e.g. BioRad, offer equipment to do that, check
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