HELP - best NOS assay

Bernard Murray bernard at elsie.nci.nih.gov
Mon Jan 8 16:11:10 EST 1996


In article <4crjv9$ggr at oac2.hsc.uth.tmc.edu>, bkone at heart.med.uth.tmc.edu 
says...
>
>Can someone please provide me with a detailed protocol for [3H]arginine 
>to [3H]citrulline conversion to measure NOS activity?
>
>Thanks.
>

A little vague a question (and I am not sure if this is indeed
the "best" assay), but...

I believe the "original" reference is;
Bredt, D.S. & Snyder, S.H. (1989) Proc. Natl. Acad. Sci. USA 86, 9030-9033

This gives a fairly detailed description of the assay.  Most other
groups use very minor modifications of this (eg. extra cofactors).
A good literature search will fill in any system-specific recommendations.
I have no idea what system you will be working with so I can't add
extra "details" to this except that my personal preference is to
include plenty of protease inhibitors when trying to assay
NOS II/iNOS in homogenates/cytosol.
	To the library...
			Bernard
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov  (National Cancer Institute, NIH, Bethesda MD, USA)




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