[ccp4bb] Offtopic: FAD enzymatic assay

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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From: Daniel Anderson dha {- at -} MBI {- dot -} UCLA {- dot -} EDU
Date: 2008-12-01

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Subject: Re: Offtopic: FAD enzymatic assay
From: conancao conan_cao {- at -} HOTMAIL {- dot -} COM
Date: 2008-12-01

Subject: Offtopic: FAD enzymatic assay
From: michael nelson gnomenm {- at -} YAHOO {- dot -} COM
Date: 2008-12-01

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

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Program to fill unitcell randomly
From: Daniel Anderson dha {- at -} MBI {- dot -} UCLA {- dot -} EDU
Date: 2008-12-01

Next message:
Subject: Re: Offtopic: FAD enzymatic assay
From: conancao conan_cao {- at -} HOTMAIL {- dot -} COM
Date: 2008-12-01