Hi colleagues, I tried to browse the archive to find the answer to this question, which I think might have been addressed before, but without success. Does anyone have a protocol for fc-receptor blocking with human IgG when staining ficoll-separated PBMCs? I usually work with 0.5 to 1 x 10 to the 6 cells per tube. My routine conjugates are CD3-APC, CD4Pe or FITC and CD8Cy-chrome. Thank you, Marcos da Silva Pediatric Immunology lab - UMass Medical School Marcos.DaSilva@umassmed.edu If you are studying t cells population in peripheral blood, you can just stain w/ CD3/4/8. These antibodies should not give you high background from nonspecific binding/fc receptor binding. I always use them and I never block fc receptor first. But, anyway, if you need to do the background control, you can do the IgG binding first before you do the staining, do it the same way you do regular staining.
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