[Protein-analysis] Re: What is interfering with SDS-PAGE?
blackhole at abuse.plus.com
Fri May 26 04:36:33 EST 2006
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
>I am solubilizing difficult proteins using a pretty harsh extraction solution:
>0.05 M DTT
>1% N-lauroylsarcosine (detergent)
>protease inbitor cocktail
>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
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.
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
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