Cd8b Mouse Monoclonal Antibody [Clone ID: 3.4.1]

CAT#: SM260RX

Cd8b mouse monoclonal antibody, clone 3.4.1, PE

Product Images

Cell Source: Thymus. Percentage of cells stained above control: 92.2%

Specifications

Product Data

• Clone Name: 3.4.1
• Applications: FC
• Recommended Dilution: Flow Cytometry, Immunoprecipitation and Western blotting.
  
  Test Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Rat Strain: Wistar.
  Cell Concentration: 1x10e6 cells per tests.
  Antibody Concentration Used: 1.0 μg/10e6 cells.
  Isotypic Control: PE Mouse IgG1, k.
  Cell Source : Percentage of cells stained above control:
  Thymus : 92.2%
  Spleen Cells : 14.6%
  Lymph Nodes : 40.3%
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Rat/mouse T cell hybrids expressing CD8.
• Specificity: This monoclonal antibody reacts with the beta chain of the CD8 differentiation antigen.
  The 3.4.1 antibody also blocks both activation in an allogenic response and cell mediated cytotoxicity by
  CD8 T cells.
• Formulation: PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE
  State: Liquid purified IgG fraction.
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography of Ascites fluid.
• Conjugation: PE
• Gene Name: Rattus norvegicus CD8b molecule (Cd8b)
• Database Link:
  Entrez Gene 24931 Rat
• Background: CD8b is expressed on most thymocytes and mature T cytotoxic/suppressor cells (MHC class I restricted). While the CD8a and CD8b form a heterodimer on the surface of thymocytes and thymus-dependent T cytotoxic/suppressor cells, the majority of NK cells, many CD8 T cells from athymic rats, many activated CD4 T cells, and intestinal epithelium lymphocytes (IEL) express CD8a without CD8b.
  This suggests that expression of the CD8 heterodimer (a/b) is more dependant on intrathymic T cell maturation than that of the homodimer (a/a). The thymus dependence of CD8a/b T cells may be due to a requirement for thymic selection on self MHC class I antigens.
• Synonyms: CD8B, CD8B1
• Note: Protocol: Flow Cytometry Analaysis:
  
  Method:
  1. Prepare a cell suspension in media A. For cell peparations, deplete the red blood cell population with Lympholyte®-Rat 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 1x10e6 cells, representing 1 test).
  4. To each tube, add 1.0 µg* dilution of SM260RX per 10e6 cells.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate the tubes at 4°C for 30 minutes.
  (It is recommended that the tubes are protected 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 propidium iodide 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 (100 μl of 2M sodium azide in 100 mls).