ATP EXTRACTION; ATPASE INHIBITORS

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!

Marcin

Martin wrote:


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.

Thanks a lot! I am doing according to what you have proposed( simple
cell lysate and it works ,I study ATP amount in cells with
luciferin-luciferase assay),
kindly regards!
Marcin