The Mouse T Lymphocyte Subset Antibody Cocktail is a three-color reagent designed to identify major subsets of T lymphocytes by direct immunofluorescent staining with flow cytometric analysis. This cocktail consists of the following antibody mixture: PE-Cy7 hamster anti-mouse CD3e (clone 145-2C11), PE rat anti-mouse CD4 (clone RM4-5), and APC rat anti-mouse CD8a (clone 53-6.7).The 145-2C11 antibody reacts with the 25-kDa ε chain of the T-cell receptor-associated CD3 complex, which is expressed on thymocytes, mature T lymphocytes, and NK-T cells of all mouse strains tested. The RM4-5 antibody recognizes CD4 (L3T4), a differentiation antigen expressed on most thymocytes, subpopulations of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T-helper cells and immunosuppressive regulatory T cells), and a subset of NK-T cells in all mouse strains tested. CD4 has also been detected on pluripotent hematopoietic stem cells, bone-marrow myeloid and B-lymphocyte precursors, intrathymic lymphoid precursors, and a subset of splenic dendritic cells. The 53-6.7 antibody reacts with the 38-kDa α and 34-kDa α' chains of the CD8 differentiation antigen (Ly-2 or Lyt-2) expressed on most thymocytes, subpopulations of mature T lymphocytes (including MHC class I-restricted T suppressor/cytotoxic cells and subsets of γδ TCR-bearing cells and intestinal intraepithelial lymphocytes) of all mouse strains tested. CD8a is also expressed on a subset of dendritic cells. The three antibodies have been titrated and pre-diluted, mixed together, and formulated for optimal staining performance. The Mouse T Lymphocyte Subset Isotype Control contains equivalent concentrations of fluorochrome- and isotype-matched negative-control immunoglobulin.
The use of three different fluorochromes for the labeling of the three different antibodies permits the recognition of each of the three antigens on each cell in a sample. The levels of expression of the three antigens distinguish the major subpopulations of developing and peripheral T lymphocytes. Additional fluorochrome-labeled reagents may be combined with the Mouse T Lymphocyte Subset Antibody Cocktail, and the Mouse T Lymphocyte Subset Isotype Control, to further characterize T-cell subpopulations.
The Mouse T Lymphocyte Subset Isotype Control contains equivalent concentrations of fluorochrome- and isotype-matched
negative-control immunoglobulin consisting of the following: PE-Cy7 Armenian hamster IgG1, κ (clone A19-3), and PE and APC Rat IgG2a, κ (clone R35-95).