SOD assay

Eric DUFOUR Eric.Dufour at cmm.ki.se
Wed Apr 11 09:31:07 EST 2001


Cu/Zn-SOD is sensitive to cyanide but not Mn-SOD (as far as I know...)
so here can be your test for each activity but in this case you have to
inhibit cytochrome c oxidase differently I guess. Pierre is doing this
routinely in his lab, so...

"Paul S. Brookes." wrote:

> The simplest assays for SOD are based on inhibition of
> xanthine/xanthine-oxidase driven reduction of cytochrome c, monitored
> in a spec at 550nm.  Look for early papers by Fridovitch & McCord for
> precise methods, or I can e-mail you a protocol sheet in MS word.As
> for distinguishing SOD1 and 2, there are no isoform-specific assays I
> know of (other than western blots for total amounts).  Also remember
> that if you have SOD2 you probably have mitochondria in your sample,
> so their cytochrome oxidase will re-oxidize the reduced cytochrome c,
> so you must add cyanide or azide to inhibit this, but not too much, or
> you'll inhibit SOD aswell - a careful titration curve is
> needed.RegardsPSB




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