Cd8a Mouse Monoclonal Antibody [Clone ID: AD4(15)]

CAT#: CL010P

Cd8a mouse monoclonal antibody, clone AD4(15), Purified

Product Images

Cell Source: Thymus. Percentage of cells stained above control: 66.0%

Specifications

Product Data

• Clone Name: AD4(15)
• Applications: CT, FC
• Recommended Dilution: Cytotoxicity assays.
  Flow Cytometry.
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration : 1x10⁶ cells per test.
  Antibody Concentration Used: 2.0 μg /10⁶ cells.
  Isotypic Control: Mouse IgM.
  Cell Source (Percentage of cells stained above control): Spleen (10.2%), Thymus (66.0%).
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgM
• Clonality: Monoclonal
• Immunogen: C57BL/6
  Donor: B6-Ly-2a spleen
  Fusion Partner: Myeloma P3/X63-Ag8
• Specificity: Anti-mouse CD8a (Ly 2.2) monoclonal antibody reacts with a sub-population of T-lymphocytes from Mouse strains expressing the Ly-2.2 phenotype but does not react with lymphocytes from strains expressing the Ly-2.1 phenotype.
• Formulation: PBS
  State: Purified
  State: Liquid purified IgG fraction
  Preservative: 0.02% Soidium Azide
• Concentration: 1.0 mg/ml
• Purification: Euglobin precipitation
• Gene Name: Mus musculus CD8 antigen, alpha chain (Cd8a), transcript variant 1
• Database Link:
  Entrez Gene 12525 Mouse
• 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 2x10⁷ cells/ml in media A. Add 50 μl of this suspension to each tube (each tube will then contain 1x10⁶ cells, representing 1 test).
  4. To each tube, add 2.0 μg of this antibody Cat.-No CL010P.
  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 (FITC Goat anti-mouse IgM-3 (H+L)) at 1/500 dilution.
  9. Incubate the tubes at 4°C for 30-60 minutes.
  (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C in media B.
  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. 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).