still waiting for suggestions please....

Chris LaRosa clarosa at biocomp.unl.edu
Sat Jul 22 10:55:02 EST 2000



"Frank O. Fackelmayer" wrote:
> 
> I partially agree with Chris and Han, but I don´t feel that the original

I would suggest that it is ok to freeze the cells in liquid Nitrogen and
then to store them for the enzymatic assays.   it may be ok to pulverize
them and then keep them frozen at -70.  However , it may depend on the
enzyme in question.    Pulverizing introduces a certain amount of
release of proteases.     Protease inhibitors are definitely called for
in the extraction buffer,,, if these do not inhibit the enzyme
reactions.    Perhaps it was a bit rude of me to rebuff the questioner,
but I stick to my opinion that the questions are too specialized and off
topic for this newsgroup.   (Molecular biology methods reagents).    the
question is more relevent in an enzymology or plant physiology
newsgroup.   Plant enzyme assays present special problems... grinding
the tissues release a S%$^load of proteases, phenolic compounds and
other interfering compounds.   I would suggest that the questioner begin
with searching agricula and Biology Abstracts, and look for specific
methods used by others .    One can try a Google.com search as well as
using search engines that specialize in science... the google.com search
may be productive,,,, I have found it surprizingly good at finding
references in molecular biology, cell biology, enzymology.  

At times some folks seem to display a lazy attitude by asking questions
very important to their research here. Furthmore the opinions expressed
here are not necessarily the best course of action... The quality of
answers should be considered very suspect and subject to further
verification.






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