This anti-mouse CD8a antigen monoclonal antibody recognizes the mouse CD8a chain. The α chain of CD8 associates with the CD8b chain to form a CD8a/b heterodimer that is expressed by the majority of thymocytes and by the MHC class I restricted subset of mature T cells1. Mouse CD8a can also form a CD8a/a chain homodimer on subsets of CD8 positive T cells. For this reason, antibodies specific for CD8a rather than CD8b are recommended for a rigorous delineation of CD8 positive cells.
Flow Cytometry Analysis (see Protocols). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
实验流程
FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test). 4. To each tube, add ~0. 5 µg* of this Ab. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. (It is recommended that the tubes areprotected from light, since most fluorochromes are light sensitive. )7. Wash 2 times at 4°C. 8. Resuspend the cell pellet in 50 µl ice cold media B. 9. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results: Mouse Strain: BALB/cCell Concentration: 1 x 10e6 cells per testAntibody Concentration Used: 0. 5 μg/10e6 cellsIsotypic Control: APC Rat IgG2a
限制
仅限研究用
浓度
0.2 mg/mL
缓冲液
0.5 mLPBS and 0.1 % sodium azide (NaN3). A highly purified grade of BSA has been added as a stabilizing protein to bring the final protein concentration to 4-5 mg/mL after conjugation.
储存液
Sodium azide
注意事项
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
储存条件
4 °C
储存方法
Store the antibody undiluted at 2-8 °C. DO NOT FREEZE!
The CD8 antigen is a cell surface glycoprotein found on most cytotoxic T lymphocytes that mediates efficient cell to cell interactions within the immune system. The CD8 antigen, acting as a coreceptor, and the T cell receptor on the T lymphocyte recognize antigen displayed by an antigen presenting cell (APC) in the context of class I MHC molecules. The functional coreceptor is either a homodimer composed of two alpha chains, or a heterodimer composed of one alpha and one beta chain. Both alpha and beta chains share significant homology to immunoglobulin variable light chains.Synonyms: CD8 alpha chain, CD8A, MAL, T-cell surface glycoprotein CD8 alpha chain, T-lymphocyte differentiation antigen T8/Leu-2