You might be right. I can not rule out the possibility.
By the way, is the PMS/DCPIP system fairly stable in the aerobic condition?
Mike
--- On Thu, 12/4/08, [EMAIL PROTECTED] <[EMAIL PROTECTED]> wrote:
From: [EMAIL PROTECTED] <[EMAIL PROTECTED]>
Subject: Re: [ccp4bb] Offtopic: F
Dear all,
I may sound stupid enough.
I tried the PMS-DCPIP assay system as suggested, and I choose to observe the
absorption at 600nM. However, when I initialize the reaction, I actually see
the Abs slowly but steadily increasing rather than decreasing.
How the FAD interact with the enzyme is
Does the FAD actually dissociate on each turnover, or does it remain bound
and transfer electrons to an acceptor? Succinate dehydrogenase is an
example of the latter, and it can be readily assayed using a mediater
phenazine methosulfate to accept the electrons and transfer them to
a redox dye dich
Hi Michael,
Fraser et al writes that in case of Synechococcus phytoene desaturase
'NAD+ and NADP+ were observed to be involved, whilst FAD was an
ineffective electron acceptor '
Biochem J. 1993 May 1;291 ( Pt 3):687-92
HTH
Guenter
PS The enzyme itself has no flavin bound?
michael nelson wro
-0800From: [EMAIL PROTECTED]: [ccp4bb] Offtopic:
FAD enzymatic assay: a little bit more about my enzymeTo: CCP4BB@JISCMAIL.AC.UK
Dear all,Thank you for all your kind replies.Here is a little bit more about
the enzyme and how I carry out the assay at the first place.My enzyme is a
lipid desaturase
Dear all,
Thank you for all your kind replies.
Here is a little bit more about the enzyme and how I carry out the assay at the
first place.
My enzyme is a lipid desaturase, originally from plant but overexpressed in
bacteria. FAD serves as a co-factor for this enzyme, in which FAD is reduced t
Mike:
The trick may be doing the assay under anaerobic condition, especially if
the FAD cofactor is sensitive to oxygen.
You need an anaerobic train and tanometers for the experiment.
Good refs: Hille, R. Biochemistry 1991 Sep 3;30(35):8522-9. Electron
transfer within xanth
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. B
