Guanidine samples on SDS PAGE

Mark Bowen mark.bowen at stanford.edu
Wed Apr 4 03:27:11 EST 2001


SDS is not soluble in Gnd at high concntrations.  you can dilute the samples if
the protein concentration is high enough.  normally I use TCA precipitation from
my Gnd solution followed by acetone rinsing and resuspension in SDS buffer with
0.1M tris pH 8.  this can be done on fairly small samples for PAGE.

Squid wrote:

> Is there some trick to this?  When I add my SDS loading buffer to the sample
> containing guanidine (6M) it produces large amounts of blue solid....I'm
> sure I should have paid more attention in chemistry class!!  My protein is
> 6XHis tagged, and must be solubilized in 6M guanidine HCl so that it can
> bind the affinity column.  I want to run the samples on a gel (quickly!!) to
> monitor purification, and was hoping not to have to dialyze the guanidine
> out first, which causes my protein to become insoluble again...
>
> thanks
>
> julie

--
Mark Bowen

Laboratory of Axel Brunger
Howard Hughes Medical Institute
Stanford University

lab (650) 736-1715
fax (650) 736-1961
mark.bowen at stanford.edu





More information about the Methods mailing list