Low CD4+ signal in functional suppression assay. Could cell death be a reason or weak staining??

05 August 2021 0 6K Report

Currently I'm performing functional suppression assays with coculturing Tregs (CD4+YFP+) and Tcon (CD4+)cells for 72 hrs. I see very weak signal of CD4+ post harvesting. What could be the reason? I therefore also cant see nice proliferative peaks.

I'm doing live dead staining with blue fluorescent reactive, is it right to do that or I should use something else to gate around my live population?

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