Hi Candace, regarding your question in how to identify gd T cells in mouse spleen. We have done quite extensive analysis of these cells in the mouse using our 10 color system. But there should be no problem in identifying the cells with 3 colors. The main point is to collect enough events (about 1% of T cells in the spleen are gd+, i.e. you want to aquire about 50.000 cells to look at a decent size population. In my hands by far the most gd T cells in mouse spleens (BALB/c, C57BL/6 and B10.BR) are CD4- CD8- and CD3+. If you use reagents from Pharmingen, you could use the CD3 Cy5PE to identify T cells (all gd T cells are CD3 positive). You will see the gd T cells as a small fraction of CD3+ cells when using the anti-gd T cell receptor antibody coupled to PE. In addition, you could put CD4 and CD8 both on FITC, gate on the CD3+, CD4/8- negative cells and then you see the gdTCR+ (PE) cells very clearly. Hope this helps, Nicole Baumgarth Stanford University School of Medicine Dept Genetics 300 Pasteur Drive, Beckman Center B007 Stanford, California 94305 - 5318 FAX: 650 - 725 8564 Phone: 650 - 723 7149
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