I am looking to measure NADH production at 340 nm by GAPDH (from crude protein extract) over time in a UV transparent 96-well plate. So far I am unable to detect any signal and I was wondering if anyone might have some tips for such an experiment.
Well in the end I used a diaphorase/rezasurin coupling reaction to shift my signal to a higher (non UV) wavelength. I did manage to obtain a signal at 340 nm but i needed very large quantities of protein extract. The advantage of the diaphorase reaction is that it is far more sensitive (less protein required).