HELP - best NOS assay
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
>Can someone please provide me with a detailed protocol for [3H]arginine
>to [3H]citrulline conversion to measure NOS activity?
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 Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
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