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

CAT#: CL058B

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

Product Images

Tissue distribution by flow cytometry analysis: Cell source spleen, percentage of cells stained above control = 99.2%

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 reacts with 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.
  Results by Flow Cytometry Analysis:
  -Tissue distribution
  Mouse Strain: C57BL/6
  Cell concentration: 1x10e6 cells per test
  Antibody concentration used: 0.1 µg/10e6 cells
  Isotypic control: Biotin Mouse IgG2a,k
  Cell source thymus: Percentage of cells stained above control = 35.5%
  Cell source spleen: Percentage of cells stained above control = 99.2%
  - Strain distribution
  Cell concentration : 1x10e6 cells per test
  Antibody concentration used: 0.1 µg/10e6 cells
  Strains tested: C57BL/6, BALB/c, C3H/He, CBA/J
  Positive: C57BL/6
  Negative: BALB/c, C3H/He, CBA/J
• Formulation: PBS containing 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid purified Ig fraction.
• Concentration: 0.1 mg IgG /ml
• Purification: Affinity Chromatography on Protein A.
• Conjugation: Biotin
• Synonyms: HLA Class I
• Note: Protocol: FLOW CYTOMETRY ANALYSIS
  Method:
  1. Prepare a cell suspension in media A. For spleen 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 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 0.1 µg of CL058B 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 appropriate 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).