Historians believe that in newspost
<mailman.1085.1148492571.16885.proteins at net.bio.net> on Wed, 24 May
2006, Rafael Garcia <rgarcia at errc.ars.usda.gov> penned the following
literary masterpiece:
>I am solubilizing difficult proteins using a pretty harsh extraction solution:
>7M Urea
>2M Thiourea
>0.05 M DTT
>0.01M Tris
>2.5% SDS
>1% N-lauroylsarcosine (detergent)
>protease inbitor cocktail
>pH7.4
>>Something in this mixture makes SDS-PAGE gels look streaky and dark. If I clean up and concentrate the extracted protein with ultrafiltration, the
>gels work much better. (dialysis followed by lyophilization doesn't seem to work). Which component in this extraction solution is causing the
>problem?
Remove one component at a time, add the remainder to any protein and
load into a single lane of a gel. Repeat removing another component etc.
Identify which component causes the problem.
Simple fault finding experiment.
Duncan
--
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
Duncan Clark
GeneSys Ltd.
