I am planning to stain lymphocytes on surface and intracellularly in whole blood. What's the better timing of RBC lysing? Before or after surface staining?
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Actually, i am more keen on the difference between RBC lysing before and after surface staining, whether and how they affect later intracellular staining et al. Do you have any experience on this aspect?
I incubate first with the extracellular antibodies (20-30min). Then, I add the RBC lysis buffer and the incubation is for 10 min. After this, I add the permeabilization/fixation buffer, and continue with the intracellular staining.