I will perform an immunoassay to detect the production of iNOS in macrophages by flow cytometry. I need to know what concentration of surfactant I should use to permeate the plasma membrane. Does anyone know of any protocol?
Haven't done it myself but seems there are a bunch of protocols out there. The easiest would be to use pre-made reagents from BD. See the technical sheet below for eNOS flow cytometry. You could use the same protocol but with an iNOS antibody instead.
http://www.bdbiosciences.com/ds/pm/tds/560103.pdf
Why are not using Griess assay promega, it is very easy to detect the concentration of nitric oxide as end product of iNOS expression by the treated cells
Hi,
I had used Griess asao for its determinaron and it works very fine.
As for intravital iNOS detection by flow cytometry, DAF-based indicators are pretty cool: https://www.thermofisher.com/cz/en/home/references/molecular-probes-the-handbook/probes-for-reactive-oxygen-species-including-nitric-oxide/probes-for-nitric-oxide-research.html#head4.
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