I'd like to look at the effect of a drug on NK and NKT cell populations. I read that one of the best markers is CD1d. But, would it be possible to get a clean gating by using CD19 to get read to get out B cells, and then use NK1.1(CD161) vs CD49b to look at NK cells? what about using NK1.1 and TCRbeta for gating NKT cells? wold it give a clean population or there'd be some non-NK or non-NKT cells among?
Thanks in advance
Hello,
NK1.1 is good for B6 but not all mouse strains express NK1.1 (e.g. Balb/c). eBioscience gives some good alternatives (Copy): The PK136 monoclonal antibody reacts with mouse NK1.1, an antigen expressed by natural killer cells and a subset of T cells in the NK1.1 mouse strains including C57BL and NZB. Several commonly used laboratory mouse strains such as BALB/c, SJL, AKR, CBA, C3H and A do not express the NK1.1 antigen. For detection of NK cells in these strains the monoclonal antibody DX5 (Cat. No. 14-5971) should be used. Simultaneous staining of C57BL/6 spleen cells with PK136 and DX5 reveals coexpression of both markers by a majority of cells as well as presence of small populations of DX5+PK136- and DX5-PK136+ cells.
With regard to NKTs, at least type I NKT cells express an invariant TCRa chain (Va14). TCRb staining in combination with CD4 and CD8 (to exclude common TCRab cells) should enrich quite well for NKTs in general and should be quite clean. You could contemplate a double stain with anti CD3 to make gating easier.
Good luck
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