Is it a must to use FC block before FACS or just a preference?
I am still doing antibodies titration and I would like to make it cheaper and less time consuming as possible.
Thank you
It depends on the cell population you are staining. Not all cell types express Fc receptor and, therefore, it would not be necessary. If you're staining PBMC, it is absolutely necessary.
If you're staining lines, it might be worth doing a pilot test with fluorophore-conjugated anti-CD16/32 to see if Fc gamma receptor is expressed on your cells if interest.
It depends on the cell population you are staining. Not all cell types express Fc receptor and, therefore, it would not be necessary. If you're staining PBMC, it is absolutely necessary.
If you're staining lines, it might be worth doing a pilot test with fluorophore-conjugated anti-CD16/32 to see if Fc gamma receptor is expressed on your cells if interest.
I almost agree with Govinda answer. But it also depends on which cells you are looking for among PBMCs (If you are going to stain PBMCs). If you desire to isolate Mo, NK cells and B cells which express Fc receptors, you definitely need blocking but T cells generally don't express Fc receptors so you can forget Fc blocking or use 5% FCS/BSA/Host serum buffer.
Hope to be helpful.
But if you're staining for T cells in a bulk PBMC population, you will falsely select some non T cells if you don't use the Fc block.
I agree with Govinda. It is possible to falsely select non-T cells if you don't use FC blocker, because paucity of B cells do express CD3. 5 or 10uL of FC works perfectly in my staining protocol.
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