b-gal assay

[Brad Nicholson]

In article <42k8gs$1m7 at eldborg.rhi.hi.is>, eiriksig at rhi.hi.is (Eirikur 
Sigurdsson) wrote:

> Hi.
 >I am having a problem doing my b-galactosidase assay.  
>snip<

Hi Eirikur,
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
===========================================================================
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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