MHC Class I H-2 Kb / H-2 Db Mouse Monoclonal Antibody [Clone ID: 5041.16.1]

CAT#: CL058R

MHC Class I H-2 Kb / H-2 Db mouse monoclonal antibody, clone 5041.16.1, PE

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Specifications

Product Data

• Clone Name: 5041.16.1
• Applications: FC
• Recommended Dilution: Flow cytometry (see protocol below).
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Specificity: This antibody is specific for cells expressing the H-2K antigen coded for by the b haplotype and for cells expressing the H-2D antigen coded for by the b haplotype.
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE
  State: Liquid Ig fraction
• Concentration: 0.1 mg/ml
• Purification: Protein G chromatography
• Conjugation: PE
• Synonyms: HLA Class I
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cell population.
  2. Wash 2 times.
  3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test).
  4. To each tube add 0.2- 0.5 mg of 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 phosphate buffered saline. (This stains dead cells by intercalating DNA.)
  
  MEDIA:
  A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 µl of 2 M sodium azide in 100 mls).
  B. Phosphate buffered saline (pH 7.2) + 0.5% bovine serum albumin + sodium azide (100 µl of 2 M sodium azide in 100 mls).
  
  FLOW CYTOMETRIC ANALYSIS:
  Donor: C57BL/6
  Cell Concentration: 1x10e6 cells
  Antibody Concentration: 0.2 µg/10e6 cells
  Isotypic Control: PE Mouse IgG2a, k
  
  (see picture below)
  
  STRAIN DISTRIBUTION:
  Procedure: As above
  Antibody Concentration: 0.2µg/10e6 cells
  Strains Tested:
  Strain Haplotype +/-
  BALB/c H-2d -
  C3H/He H-2k -
  CBA/J H-2k -
  C57BL/6 H-2b +
  B6Lyt 2.1 3.1 H-2b +