SDS-problems

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Wed Oct 20 11:47:56 EST 1999


In bionet.molbio.methds-reagnts Pedro Silva <Pedro.Silva at nospamepr.bc.wau.nl> wrote:
> I have partially purified a membrane protein from a hyperthermophilic
> organism. The protein has been solubilized with 0.4% DOC, then
> precipitated w/ 15% ammonium sulfate and resuspended in 0.4% DOC
> -containing buffer. 
> Then I tried to run a SDS-Phast Gel with it, but the protein did not
> migrate at all. I had boiled it for 10 min w/ SDS and
> beta-mercaptoethanol.

Membrane proteins often have the nasty property of forming large
insoluble aggregates when being boiled in the presence of SDS. Try
incubating it in 5x Laemmli at approx. 30°C for a while.

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */



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