While performing intracellular cytokine assay on B cells, I am finding a peculiar population. I use 2*10^6 cells for studying each functional marker. The pbmcs are cultured for 72 hours, with CD40L dose being given in 0 hour after plating and BFA in the 48th hour. When analysed, I am getting two CD19+ populations from the lymphocytes. Can anyone enlighten me about what actually is happening?

  • $\begingroup$ Have you phenotyped the populations further, other than using CD19 after presumably gating on CD3-? How are you detecting the two populations of CD19+? Are you plotting CD19 vs another marker, or is it simply CD19 intensity? Can you share your plots and gating strategy, comparing untreated vs treated? Also, what is BFA - brefeldin A? $\endgroup$
    – MattDMo
    Mar 18 at 1:15

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