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

CAT#: CL010F

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

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Specifications

Product Data

• Clone Name: AD4(15)
• Applications: FC
• Recommended Dilution: Cytotoxicity assays.
  Flow Cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgM
• Clonality: Monoclonal
• Immunogen: C57BL/6
  Donor: B6-Ly-2a
  Fusion Partner: Myeloma line P3/X63Ag8
• Specificity: Anti-mouse CD8a (Ly 2.2) monoclonal antibody reacts with a subpopulation 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 containing 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: FITC
  State: Liquid purified Ig fraction
  Label: Fluorescein isothiocyanate isomer 1
• Purification: Euglobin precipitation
• Conjugation: FITC
• 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 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 2.0-1.0 μg of this antibody 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.
  (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).
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration : 1x10e6 cells per tests
  Antibody Concentration Used: 2.0 μg/10e6 cells
  Isotypic Control: FITC Mouse IgM
  Percentage of cells stained above control:
  Spleen 10.2%
  Thymus 66.0%