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Two(ish) questions about NK cells and/or FACS

TG
in Microbiology and Immunology

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I'm new here. wot.

 

1. I was wondering what would be the best way to quantify the number of NK cells in a tumor. The tumors would be in SCID mice. The problem is that the percentage of NK cells is pretty low, so when I FACS the cells, I might need to back-gate PE+ cells (the cells are stained with DX5-PE) in order to increase the resolution, and get quantifiable quantities.

 

2. I was also wondering if any of you guys know what the percentage of NK cells in a mouse spleen is... Specifically, in BALB/C SCID mice and in WT B6 mice, but any info would help. I did the googles, but got some conflicting results. I need this as a control for my back-gating, so if backgating is a bad idea, nevermind.

 

Bonus question: is it ok to quantify NKs using one antibody (DX5), or should I have two antibodies? (and if so, which?) Also, I think DX5 stains NKT cells too, but since I'm working on SCID mice, I doubt they have NKT cells, so this shouldn't be a problem. I think.

 

helps. thnxs.

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