What is the reason for not getting nitric oxide result in spleenocytes stimalated with LPS?

15 March 2023 0 355 Report

I stimulated the isolated splenocytes with LPS (1 ug) and incubated them for two different time points 24hrs and 48 hrs. Then checked the Nitrite levels in the culture supernatant were by using Griess reagent (0.1% sulfanilamide in 3% phosphoric acid and 1% NED). But I didn't get any results, even though I used a higher concentration of LPS (10 ug) and incubated for longer (72 hr). When I used the same LPS and treated RAW macrophages, I got the result. Can anyone explain the correct protocol for nitric oxide estimation in splenocytes upon stimulation with LPS?

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