was working to set up an FAD enzymatic assay. I wished to be able to use 450nM
to continuously monitor the progress of the reaction. The substrate I used is
the natural substrate of the enzyme and the protein is recombinant protein and
I assume it's active since I do see changes in TLC plate. But no signal was
observed at all using a spectrometer. Does anyone here have any suggestions on
how to correctly carry out FAD enzymatic assay?
Also I heard from one postdoc here that FADH2 is quickly reoxidized by O2. Is
that right? Should I do the assay under anaerobic condition? If so, how?
Your help is highly appreciated!
Thanks!
Best regards,
Mike
