I am using ImmunoCult™ Human CD3/CD28 T Cell Activator (Semcell, Catalog #10971) to activate PBMC and expand T cell. but the T cell activator contains an anti-CD3 antibody clone that fully or partially blocks all anti-CD3 antibody clones used to assess CD3 expression by flow cytometry.
Are there any other ways to avoid the interference of CD3 in the CD3/CD28 T Cell Activator it I have to gate CD3 by FACS? Is it possible to avoid this if I use a different clone No. of CD3 for FACS analysis?
I'm not sure that this can be avoided. I would try using a CD4 and CD8 antibody to gate on. This way you get both CD4 and CD8, but avoid the CD3 issue as stated above. If you're sorting then you could put both CD8 and CD4 in the same fluorophore and you should end up with all the T cells.
Have your tried to add anti-CD3 antibody as an intracellular stain? Sometimes during activation, CD3 down regulates and this effects the gating. Many people add CD3 as a part of their intracellular stain to avoid this. This is applicable if you need to analyse the data but not for sorting
Jessica M. Reel Thanks a lot! It appears this cannot be avoided, I'll try CD4 and CD8 or others instead.
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