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ATP EXTRACTION; ATPASE INHIBITORS

ATP EXTRACTION; ATPASE INHIBITORS - Protocols and Methods Forum

ATP EXTRACTION; ATPASE INHIBITORS - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 05-06-2004, 11:03 AM
Martin
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Default 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
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  #2  
Old 05-10-2004, 05:23 AM
Dr Engelbert Buxbaum
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Default ATP EXTRACTION; ATPASE INHIBITORS

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.
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  #3  
Old 05-10-2004, 02:23 PM
Martin
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Default ATP EXTRACTION; ATPASE INHIBITORS

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
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