ATP EXTRACTION; ATPASE INHIBITORS

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Mon May 10 00:23:26 EST 2004


Martin wrote:

> Hey,
> 
> I will be really gratefull if somebody tell how extract ATP from human
> cells.
> There is also another problem with ATPases - so after extraction we
> should use ATPases inhibitors.
> Maybe somebody knows which inhibitors are posible to use?
> BEST WISHES!

ATPases first: Plasma membrane Na/K-ATPase is inhibited specifically
with ouabain (0.1 mM), F-type ATPases with NaN3 (3 mM). Depending on
what you want to do, vanadate can inhibit quite a few ATPases, but is
difficult to get rid of later. You may also wish to work at a slightly
acidic pH.

Isolation method depends on what you want to do: preparative isolation
of ATP (forget it: yeast is a much cheaper source) or determination of
ATP-concentration, e.g. with luciferase. In the latter case, no
isolation is actually required, a simple cell lysate will do.



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