HLA Class II DR Rat Monoclonal Antibody [Clone ID: YD1/63.4.10]

CAT#: SM2201R

HLA Class II DR rat monoclonal antibody, clone YD1/63.4.10, PE

Product Images

Cell Source: Peripheral Blood Lymphocytes

Specifications

Product Data

• Clone Name: YD1/63.4.10
• Applications: FC
• Recommended Dilution: Flow Cytometry.
  Immunohistochemistry on Cryostat Sections.
  Immunohistochemistry on Paraffin-Embedded Sections.
• Reactivities: Human
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: DAUDI cells. Donor: immunized DA rat spleen cells. Fusion Partner: Y3 Ag1.2.3 rat myeloma
• Specificity: This HLA-DR monoclonal antibody recognizes the HLA-DR (MHC class II) antigen.
• Formulation: PBS
  Label: PE
  State: Liquid purified Ig fraction from bioreactor supernatant
  Stabilizer: EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Preservative: 0.02% Sodium Azide
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: PE
• Synonyms: HLA-DR, HLA class II histocompatibility antigen DR, MHC class II antigen DR
• 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®-H 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 1 x 10⁶ cells, representing 1 test).
  4. To each tube, add 1.0–0.5 μg* of this antibody per 10⁶ 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 be 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:
  Cell Concentration: 1x10⁶ cells per test.
  Antibody Concentration Used: 0.5 μg/10⁶ cells.
  Isotypic Control: PE Rat IgG2a.