BD, 557916, BD Pharmingen™ Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control

Reactivity: Mouse (QC Testing)
Application: Flow cytometry (Routinely Tested)
Regulatory Status: RUO
RRID: AB_396937
Description: Description The Mouse T Lymphocyte Activation Antibody Cocktail is a three-color reagent designed to identify major subsets of T lymphocytes by direct immunofluorscent staining using flow cytometric analysis.  This cocktail consists of the following antibody mixture:  PE-Cy™7 rat anti-mouse CD25 (clone PC61), PE Armenian hamster anti-mouse CD69 (clone H1.2F3), and FITC Armenian hamster anti-mouse CD3ε (clone 145-2C11).  PE-Cy™7 rat anti-mouse CD25 (clone PC61) reacts with CD25, the low-affinity IL-2 receptor α chain (IL-2Rα, p55) expressed on activated mouse T and B lymphocytes.  CD25 is also found on some developing B cells in the bone marrow, early developing T cells in the thymus, peripheral CD4+ regulatory T (Treg) cells, and dendritic cells.  PE hamster anti-mouse CD69 (clone H1.2F3) reacts with CD69, also known as the very early activation antigen.  Its expression is rapidly induced upon activation of lymphocytes (T, B, NK, and NK-T cells), neutrophils, and macrophages.  CD69 has also been reported to be expressed on thymocytes that are undergoing positive selection.  FITC hamster anti-mouse CD3ε (clone 145-2C11) reacts with the 25 kDa ε chain of the T-cell receptor-associated CD3 complex, expressed on thymocytes, mature T lymphocytes, and NK-T cells. The Mouse T Lymphocyte Activation Isotype Control contains equivalent concentrations of fluorochrome- and isotype-matched negative-control immunoglobulin consisting of the following: PE-Cy™7 rat IgG1, λ (clone A110-1), PE Armenian hamster IgG1, λ (clone G235-2356) and FITC Armenian hamster IgG1, κ (clone A19-3).
Preparation And Storage: Preparation And Storage Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. The antibody was conjugated with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
Recommended Assay Procedures: Recommended Assay Procedures Flow cytometry: The three antibodies contained within the Mouse T Lymphocyte Activation Antibody Cocktail have been titrated, pre-diluted, mixed together, and formulated for optimal staining performance.  The use of three different fluorochromes for labeling of three different antibodies allows for the distinct recognition of three different 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 Activation Antibody Cocktail, and the Mouse T Lymphocyte Activation Isotype Control, to further characterize T-cell subpopulations.