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NAD+ and DTT


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#1 oscartirado

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Posted 06 May 2010 - 06:19 AM

I'm doing an enzyme assay where I measure reduction of NAD+ at 340nm. I do this with cell free extract in 50mM phosphate buffer and 1mM DTT. My question is: will DTT interfere with my assay (can DTT reduce NAD+)?

Thanks,
Oscar

#2 mdfenko

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Posted 06 May 2010 - 10:34 AM

i used to perform assays that reduced nad (and nadp) with enzyme in solution with 2-mercaptoethanol. there was no effect on the reduction.

now that i think of it, i also performed nad reducing assays in the presence of dtt and there appeared to be no effect.

(the final concentration of reducing agent in the assay was low, sub mM for dtt)

Edited by mdfenko, 06 May 2010 - 10:34 AM.

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