Temp. staining of SDS PAGE gels prior to renaturation??
mack at fcs260c.ncifcrf.gov
Sat Nov 13 23:01:10 EST 1993
In article <CGFpD7.ELs at usenet.ucs.indiana.edu> Bill Nowatzke <wnowatzk at ucs.indiana.edu> writes:
>I have been trying to follow a R. Burgess paper in which enzymatic
>activity was regained in a protein purification after crushing the
>suspected protein band from the gel. The problem that I have been having
>is the transient staing of the gel to identify the approx Mr prior to
>cutting out the band. The original paper recommends ppt the SDS in the
>gel with 0.2M KCl for 5 min and then destaining with water for up to one
>hr. When I tried this the portion of the gel below the dye front became
>very white, implying ppt, but the portion of the gel above the dye front
>only became slight opaque and I could only see very faint bands of heavly
>overloaded lanes. I was made aware of another transient staining tech
>using a 0.3M copper soln which I tried on the same gel with poor results
>(although this gel would not fully destaing from the KCl treatment and
>that probably interfered).
>I would appreciate any advice/procedures that you may have successfully
>used or heard about. Thanks in advance!
I've heard of people soaking the gel in KOAc, where the protein becomes
opaque. Joe Mack mack at ncifcrf.gov
More information about the Proteins