MHC Class II I-Ab Mouse Monoclonal Antibody [Clone ID: 25-5-16S]

CAT#: SM080FS

MHC Class II I-Ab mouse monoclonal antibody, clone 25-5-16S, FITC

Product Images

Cell Source: Spleen. Percentage of cells stained above control: 49.3%

Specifications

Product Data

• Clone Name: 25-5-16S
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgM
• Clonality: Monoclonal
• Specificity: Recognizes Mouse MHC class II I-Ab, private for I-Ab.
  This Monoclonal antibody specifically reacts with the I-Ab
  encoded MHC class II antigen expressed on Mouse strains of the H-2b haplotype.
• Formulation: PBS containing 0.02% Sodium Azide as a preservative as a and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: FITC
  State: Liquid purified IgM fraction
• Concentration: 0.1 mg/ml
• Conjugation: FITC
• Background: Class II antigens are most highly expressed on antigen-presenting cells including B cells, macrophages, dendritic cells and certain epithelial cells.
• Synonyms: H2-Eb1, H-2 class II histocompatibility antigen I-A beta chain
• 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 1x10e6 cells, representing 1 test).
  4. To each tube, add~1.0 µg of SM080FS 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 ml of 2M sodium azide in 100 mls).
  B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 ml of 2M sodium azide in 100 mls).
  
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse strain: C57BL/6
  Cell Concentration: 1x10e6 cells per test.
  Antibody Concentration Used: 1.0 µg/10e6 cells.
  Isotypic Control: FITC Mouse IgM