Cd8a Rat Monoclonal Antibody [Clone ID: YTS169.4]

CAT#: CL007B

Cd8a rat monoclonal antibody, clone YTS169.4, Biotin

Product Images

FLOW CYTOMETRY ANALYSIS Cell Source: Thymus Percentage of cells stained above control: 92.8%

Specifications

Product Data

• Clone Name: YTS169.4
• Applications: FC
• Recommended Dilution: Flow Cytometry Analysis (see Protocols).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: Murine thymocytes
  Donor: (LOU X DA) F1 rats
  Fusion Partner: Myeloma Y3/Ag 1.2.3
• Specificity: This anti-mouse CD8a (Ly 2) monoclonal antibody reacts with a protein of approximately 30 kDa found on mouse thymocytes and mouse cytotoxic/ suppressor T cells. It does not bind to mouse helper/inducer T cells. It binds to T lymphocytes from all mouse strains regardless of phenotypic expression (i.e. reacts with T lymphocytes from mouse strains expressing the Ly 2.1 or Ly 2.2 phenotype). It can be used to investigate the role of T cells in models for infectious disease, autoimmunity, transplantation tolerance and fundamental aspects of immunology.
• Formulation: PBS, 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid purified IgG fraction
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: Biotin
• Database Link:
  Entrez Gene 12525 Mouse
• Background: 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
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population 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 this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test).
  4. To each tube, add 0.1-0.2 µg* of this Ab per 10e6 cells.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate the tubes for 30 minutes at 4°C.
  7. Wash 2 times at 4°C.
  8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1:500 dilution.
  9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C.
  11. Resuspend the cell pellet in 50 µl ice cold media B.
  12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in PBS. The 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 (100 µl of 2M sodium azide in 100 mls).
  
  Results - Tissue Distribution
  Mouse Strain: BALB/c
  Cell Concentration: 1x10e6 cells per tests
  Antibody Concentration Used: 0.2 µg/10e6 cells
  Isotypic Control: Biotin Rat IgG2b
  
  Results - Strain Distribution
  Cell Concentration: 1x10e6 cells per tests
  Antibody Concentration Used: 0.2 µg/10e6 cells
  Strains Tested: BALB/c, CBA/J, C3H/He,C57BL/6
  Positive: BALB/c, CBA/J, C3H/He,C57BL/6
  Negative: none