Inactivating enzymes before [NADH] assay

Emir ekhatipo at NOSPAMmidway.uchicago.edu
Tue Nov 7 17:23:03 EST 2000


Why do you need to keep the NADH after the reaction? For a subsequent assay?
Any specific reason to do that and not to
As for inactivation, I could think of, e.g., immunoprecipitating your
enzyme, certainly if you cannot imagine of another natural inhibitor.

Emir

"Tim Barnett" <tbarnett at bimcore.emory.edu> wrote in message
news:3A0887DD.55A67663 at bimcore.emory.edu...
> Hi all,
>
> I am interested in performing a coupled NAD-dependent enzyme assay where
> I will measure the amount of NADH produced after a period of time.
> However, I need a way to inactivate the enzyme without affecting the
> NADH in the reaction mixture. It appears that NADH is sensitive to both
> acid and heat, which are the only strategies I can think of. If anyone
> has any ideas on how to do this I would be very thankful.
>
> - Tim
>
> --
> Tim Barnett, PhD
> Postdoctoral Research Fellow
> Department of Microbiology and Immunology
> Rollins Research Center
> Emory University
> 1510 Clifton Road
> Atlanta, GA 30322
>
> Ph: (404) 727 0522
>
>







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