MHC Class II RT1D Mouse Monoclonal Antibody [Clone ID: OX-17]

CAT#: CL121R

MHC Class II RT1D mouse monoclonal antibody, clone OX-17, PE

Product Images

Cell Source: Spleen. Percentage of cells stained above control: 49.6%

Specifications

Product Data

• Clone Name: OX-17
• Applications: FC
• Recommended Dilution: Flow Cytometry.
  Tissue Distribution by Flow Cytometry Analysis:
  Rat Strain: Buffalo.
  Cell Concentration : 1x10e6 cells per test.
  Antibody Concentration Used: 0.5 μg/10e6 cells.
  Isotypic Control: PE Mouse IgG1
  Cell Source: Percentage of cells stained above control
  Thymus: 13.3%
  Spleen: 49.6%
  Lymph Node: 25.4%
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Rat spleen membrane glycoproteins depleted of Ia-A antigens.
• Specificity: Recognizes Rat RT1.D (Rat Ia-E).
  This RT1.D monoclonal antibody recognizes a monomorphic determinant on the a chain of the rat Ia antigen and appears to be the Rat homologue of Mouse Ia-E. It recognizes the rat Ia product present on B, but not T cells from lymph node or thoracic duct lymph. It does not bind to thymocytes or erythrocytes.
  The antibody does not cross-react with Rat Ia-A or Mouse Ia-E antigen, but Rabbit antibody raised against the antibody affinity column-purified MRC OX-17 antigen cross-reacted on tissues of mice expressing Ia-E Mouse antigen but not on those Mouse strains that were Ia-E antigen negative (2).
• 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.
• Conjugation: PE
• Synonyms: HLA Class II
• 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®-Rat Cell Separation Medium (CL5040).
  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 0.5 μg of CL121R 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).