Nasty NADH Nightmare
M.Purnell at botany.uq.edu.au
Fri Sep 11 00:03:07 EST 1998
I am having trouble preparing my NADH solutions for spectrophotometric assays.
I prepare a 1 mg ml-1 solution, without vortexing, in 100mM Tris pH 8.0.
Sometimes (but not always) I find that blanking on the NADH solution and then
following the absorbance change at 340 nm doesn't give me a nice, straight
line, but rather an absolutely crazy read (it jumps all over the place). I've
ruled out machine error, always use fresh NADH, and have confirmed the pH is
8.0 (i.e., the pH is not too low).
Can anyone suggest what is going on and how to fix it?
University of Qld
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