sulfhydryl availablility

Wolfgang Schechinger hubahopp at gmx.de
Wed Jul 28 17:58:24 EST 2004


Dear Arthur, 

if you have purified protein, you might see the SH in NMR spectroscopy or
in MS fragmentation patterns. There are reagents like NBD-F (available from
SAF / Fluka or Molecular Probes, I think) for fluorescent labelling. So if
you compare WT with your mutation... and in situ labelling followd by
purification with purified protein labelled after storing in a reducing
(DTT, BME) environment or oxidative conditions (H2O2, peracids, oxygen
bubbling, ...)

Depends of course a little on how many SH groups are present per molecule
in order to have a good S/N ration, but you should find some details in the
Methods in Enzymology series in your library.

just some thoughts,

Wo

BTW what are IV curves?


At 08:15 27.07.2004 -0400, you wrote:
>Hello,
>
>I have transfected my astrocytes with Kv1.1 DNA with a 359C mutation 
>(sulfhydryl).
>
>I know that the channels express, I can see this from IV curves.
>
>I would now like to modify the sulfhydryl.
>
>Could anyone tell me if there is a definitive way to know whether the 
>-SH group is present and available for binding?
>
>Thank you,
>Arthur Beyder
>
>
Wolfgang Schechinger


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