MHC Class II I-Abd Mouse Monoclonal Antibody [Clone ID: 28-16-8S]

CAT#: CL067B

MHC Class II I-Abd mouse monoclonal antibody, clone 28-16-8S, Biotin

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Specifications

Product Data

• Clone Name: 28-16-8S
• Applications: FC
• Recommended Dilution: Flow cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgM
• Clonality: Monoclonal
• Specificity: This monoclonal antibody reacts with the I-Ab encoded MHC class II antigen expressed on mouse strains of the H-2b haplotype. It also reacts with the I-Ad encoded MHC class II antigen expressed on mouse strains of the H-2d haplotype. Class II antigens are most highly expressed on antigenpresenting cells including B cells, macrophages, dendritic cells and certain epithelial cells.
• Formulation: PBS and contains 0.02% sodium azide (NaN3) as a preservative.
  Label: Biotin
  State: Liquid purified IgM
• Concentration: 0.1 mg/ml
• Purification: Affinity chromatography
• Conjugation: Biotin
• 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 0.2µg – 0.5mg* of this Ab 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. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1/500 dilution.
  9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C.
  11. Resuspend the cell pellet in 50 µl ice cold media B.
  12. 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 test
  Antibody Concentration Used: 0.5 µg/10e6 cells
  Isotypic Control: Biotin Mouse IgM