method(s) to measure rATP
Lab_Mac_Hanson at QMrelay.mail.cornell.edu
Wed Sep 7 12:17:07 EST 1994
In article <1994Sep6.141917.1 at eagle.wesleyan.edu>,
tjames at eagle.wesleyan.edu wrote:
> I am looking for a very sensitive assay method for measuring ATP levels in
> biological samples. I cannot use radioactivity.I am not current on the
> literature in this area. Could someone give me pointers to any methods.
Way back a LOOOOONG time ago, I measured ATP levels in samples, ( and, in
fact, their specific activities) using firefly luciferase. Essentially,
one adds the crude enzyme to vials w/ nucleotides and then slams the vials
into a scintillation counter with 3H window (I think this is right).
Letting the machine count every 0.1 min or so one will get decreasing
counts (corresponding to light in the sample), and can then extrapolate to
zero time. It seems sortof crude, but it worked well (at least by late
70's standards). I can't remember where I got the technique, but it should
have been dated circa mid-70's. I had two articles in j. Supramolec.
structure in I think 1979 that might reference it. Good luck !
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