pombe cell wall

S L Forsburg forsburg at nospamsalk.edu
Tue Jun 24 09:13:59 EST 1997


> Hi,
> I want to digest the cell walls from my S. pombe. The protocols I have
> found
> use Novozyme sometimes together wil Zymolase. The problem is Novozyme
> has
> some protease activity, which is a bad thing because I want to do some
> 
> immunofluorescence on a plasma membrane protein I have expressed. Does
> 
> anyone know of any other enzymes, that have no protease activity, that
> I can
> use? Can I use Zymolase alone?

You can use Zymolyase alone.  It eats up different 
sugar  linkages than Novozyme so is rather less efficient 
than the mix.

Also, you can clean up the Novozyme a bit, by doing a sulfate cut
on it.  I've never done this myself so I don't remember where it
comes down, although I do remember that it makes a 
difference with respect to proteases.  I'll try to track  down
the conditions  and post it to the newsgroup. 

-- 
-susan
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S L Forsburg, PhD  forsburg at salk.edu
Molecular Biology and Virology Lab          
The Salk Institute, La Jolla CA 
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