Nasty NADH Nightmare

skorycd at mail.ncaur.usda.gov skorycd at mail.ncaur.usda.gov
Mon Sep 14 12:36:36 EST 1998


I'm somewhat confused.  Why are you blanking on the NADH solution.  The O.D.
can only go down with NADH(H).

    NADH(H) ----> NAD+     *results in a decreased OD 340

Perhaps knowing what type of assay you are performing will help diagnose the
problem.  Feel free to E-mail me and I'll try to help.  Be warned .. I'm no
expert. But, we do perform various dehydrogenase assays here in our lab.

Christopher D. Skory
Research Microbiologist
skorycd at mail.ncaur.usda.gov
Peoria, IL

In article <6taaub$akp$1 at bunyip.cc.uq.edu.au>,
  M.Purnell at botany.uq.edu.au (Matt Purnell) wrote:
> Hello,
>
> 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?
> Thanks.
>
> Matt Purnell
> Botany Dept.
> University of Qld
>
>

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