MHC Class II I-Ak Mouse Monoclonal Antibody [Clone ID: 14V.18]

CAT#: SM083FS

MHC Class II I-Ak mouse monoclonal antibody, clone 14V.18, FITC

Product Images

Cell Source: Spleen. Percentage of cells stained above control: 75.4%

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Specifications

Product Data

• Clone Name: 14V.18
• Applications: Assay, FC
• Recommended Dilution: Flow Cytometry (See Protocols).
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: A.TL spleen.
  Donor: A.TH spleen.
  Fusion Partner: P3-X63-Ag8
• Specificity: This Monoclonal antibody is a cytotoxic antibody specific for cells expressing the Ia antigen coded for by the A subregion of the k haplotype. The reaction pattern of this antibody with a panel of inbred and recombinant haplotypes demonstrates that the antibody reacts with Ia.m2, a private specificity of the H-2k haplotype.
  This antibody can be used to quantitate or to eliminate cells bearing the I-Ak (Ia.m2) antigen.
• 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: FITC
  State: Liquid purified IgG fraction
  Label: Fluorescein
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: FITC
• Synonyms: H2-Aa
• 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 106 cells, representing 1 test).
  4. To each tube, add 1.0 μg* of SM083FS 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: A.TL
  Cell Concentration : 1x10e6 cells per test.
  Antibody Concentration Used: 1.0 μg/106 cells.
  Isotypic Control: FITC Mouse IgG2a
  
  Cell Source Percentage of cells stained above control:
  Thymus: 32.5%
  Spleen: 75.4%
  Lymph Node: 32.1%
  Bone Marrow: 21.6%
  
  Strain Distribution by Flow Cytometry Analysis:
  
  Cell Concentration : 1x10e6 cells per test.
  Antibody Concentration Used: 1.0 μg /10e6 cells.