request for NO assay protocol

Paul S. Brookes. brookes at uab.edu
Tue Jan 25 13:48:46 EST 2000


Remember that the Greiss reaction DOES NOT MEASURE NO, only nitrate and 
nitrite, which can be made from a variety of sources including NO.

WPI sells NO electrode tips for their ISO-NO system that apparently go down 
to the low nM range but they're very fragile and of course can only measure 
NO released from cells.   You could also try using a hemoglobin to 
methemoglobin assay system (the "Methods in Nitric Oxide reasearch" book 
should have a protocol), again remembering that this will only measure NO 
that is released from the cells.

What cell type are you using?  Are they stimulated with anything (LPS, 
ifn-gamma) to turn on iNOS or are you relying on basal levels?   If it's 
human tissue homogenates, make sure there's no hemoglobin contamination, as 
that will soak up NO pretty effectively.

Regards
PSB


_________________________________________
Dr. Paul S. Brookes.            (brookes at uab.edu)
UAB Department of Pathology,   G004 Volker Hall
1670 University Blvd., Birmingham AL 35294 USA
Tel (001) 205 934 1915     Fax (001) 205 934 1775
http://peir.path.uab.edu/brookes

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