brad at howard.genetics.utah.edu
Wed Sep 6 14:31:03 EST 1995
In article <42k8gs$1m7 at eldborg.rhi.hi.is>, eiriksig at rhi.hi.is (Eirikur
>I am having a problem doing my b-galactosidase assay.
Our lab does quite a lot of b-gal assays with E. coli. From your post I
think that you are in some sort of bacteria, so my info may help.
First, we just use a drop of toluene to permeabilize E. coli. If there is
a reason that you don't want to use toluene, then you may want to look at
the July, 1995. BioTechniques. Schupp, Travis, Price, Shand & Keim. "Rapid
bacterial permeabilization reagent useful for enzyme assays." p.18-20.
They describe using polymyxin B sulfate to permeabilize cells. They also
gave graphs of the relative effectiveness when many different lysis
reagents were used. It looks like polymyxin B sulfate or "Koch's" lysis
reagent(Toluene + other goodies) gave the best results with b-gals.
Good luck, hope this helps.
Brad Nicholson |"If it worked the first time, it wouldn't be
Department of Pathology | research."...Brad Nicholson
University of Utah |
Salt Lake City, UT 84132 | iligitimi non corborundrum
brad at howard.genetics.utah.edu |
or: (801)-581-4365 | My opinions are solely my own.
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