Hello,
I am trying to measure NO in cell culture (fibroblast cells).
Using Griess reagent I try to follow the paper from Wang et al., 1996 which gives a basic protocol that does not differ from the newer protocols nowadays (using Griess reagent).
Till now, I was not able to measure any NO in the supernatant of the cells.
Has anyone experience on how to prepare the supernatant?
I tried to take it directly from the cells but I assume it was not concentrated enough. Growing the cells again, using less media I spin down the supernatant to avoid false pos. results due to cell debris. But I still could not detect any signal.
I am currently waiting for one of the 'bigger' Griess kits including an additional reductase step.
Thanks a lot for help and suggestions.
Simone
Hi Simone, just by chance I saw minutes ago a related discussion on using Griess reagent. Lots of opinion, experience and suggestions. Check this out https://www.researchgate.net/post/Nitric_Oxide_Assay2
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