Hi all, I need to can we measure i-NOS in h.pylori treated samples collected in PBS?
After infection of any cell line , we usually washed cells with PBS before collection of cell pellet for RNA extraction.
I-NOS will be released in supernatant, I wonder since it in an colorimetric method, so if we have already collected samples in PBS, can we still major same?
You can either measure NO using Griess assay (colorimetric)or measure the expression of iNOS using qPCR. I am not clear on what samples you have collected in PBS. If I understand you correctly, you removed supernatants, washed the wells with PBS and want to know if you can measure NO in the PBS wash. The answer is no. You need to collect supernatants from your H.pylori infected cells and use those for Griess assay. Alternatively, you can look at the expression of iNOS in the infected cells by doing qPCR.
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