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

CAT#: SM2201F

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

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Specifications

Product Data

• Clone Name: YD1/63.4.10
• Applications: FC
• Recommended Dilution: Flow cytometry.
  Immunohistochemistry on cryostat and 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: Anti-human HLA-DR monoclonal antibody recognizes the HLA-DR (MHC class II) antigen.
• Formulation: PBS containing 0.02% sodium azide (NaN3) as preservativeand 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
• Concentration: lot specific
• Purification: Affinity chromatography on Protein G
• Conjugation: FITC
• Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Synonyms: HLA-DR, HLA class II histocompatibility antigen DR, MHC class II antigen DR
• Note: Protocol: FLOW CYTOMETRYANALYSIS:
  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 2x10e7 cells/ml in media A. Add 50 μl of this suspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test).
  4. To each tube, add 50μl of a 0.2-0.1μg dilution 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.
  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: 1x10e6 cells per test
  Antibody Concentration Used: 0.1 μg/10e6 cells
  Isotypic Control: FITC Rat IgG2a