>> I am studying the redox properties of my enzyme using various redox
> buffers (GSH/GSSG; cysteine:cystine; DTTred/DTToxid). Most of the
> protocols that have described in the literature for these buffers
> don't
> describe whether these buffers were deoxygenated prior to use; or
> whether any precautions were taken to prevent oxidation during the
> assay. Are there any protocols for redox buffers that take the effect
> of oxygen into account ?
I usually add a bit of EDTA (preventing metal-ion catalyzed oxidation)
and purge extensively with Argon.
>> One more question: the active site cysteines of may enzyme may become
> oxidised to sulfinic or sulfenic acids ... how could I test this ?
Ellman?
Sequencing?
IEF?
Pyridine-ethylation and Amino acid analysis? ...
HTH
Kresten
>> Thank you for your time,
>> Che Pillay
> Dept. of Biochemsitry
> University of Natal
> South Africa
>pillayche at nu.ac.za>> ---
>
--
The address kresten at my-dejanews.com is for
spambots only. Please mail me at LysLeuLeu at crc.dk
transforming the pre at -part into my initials.
Kresten Lindorff Larsen, Dept. Yeast Genetics
Sent via Deja.com http://www.deja.com/
Before you buy.
