MHC Class II I-Ap Mouse Monoclonal Antibody [Clone ID: 7-16.17]

CAT#: CL072B

MHC Class II I-Ap mouse monoclonal antibody, clone 7-16.17, Biotin

Product Images

Figure 1

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  Figure 2: strain distribution

Specifications

Product Data

• Clone Name: 7-16.17
• Applications: Assay, FC
• Recommended Dilution: Flow Cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: B10.P
  Donor: BALB/c
  Fusion Partner: SP2/0
• Specificity: This monoclonal antibody is a cytotoxic antibody which defines a public I-A antigen. This antibody reacts with I-A antigen from the following I-A haplotypes: I-Ap,k,q,r,s,b. Using recombinant strains, reactivity against the b haplotype has been localized to the Ab subregion. This antibody can be used to quantitate or eliminate I-A bearing cells or for precipitating I-A antigen.
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: Biotin
  State: Liquid purified Ig
• Concentration: 0.1 mg/ml
• Purification: Protein G 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 1.0 - 0.5 µg* 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: BDP
  Cell Concentration: 1x10e6 cells per tests
  Antibody Concentration Used: 0.5 µg/10e6 cells
  Isotypic Control: Biotin Mouse IgG2a
  
  Cell Source Percentage of cells stained above control:
  Thymus: 36.5%
  Spleen: 51.5%
  Lymph Node: 16.3%
  Bone Marrow: 24.4%
  
  Strain Distribution by Flow Cytometry Analysis:
  Antibody Concentration Used: 0.5 µg/10e6 cells
  Strains Tested: see FIGURE 2; For a more detailed strain distribution - see reference 1.