MHC Class II I-Ad Mouse Monoclonal Antibody [Clone ID: 34-5-3S]

CAT#: CL090R

MHC Class II I-Ad mouse monoclonal antibody, clone 34-5-3S, PE

Product Images

Flow cytometric analysis: Cell source is spleen. Percentage of cells stained above control: 52.0%

Specifications

Product Data

• Clone Name: 34-5-3S
• Applications: FC
• Recommended Dilution: Flow cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: BDF splenocytes
• Specificity: This antibody is a cytotoxic monoclonal antibody specific for cells expressing the Ia antigen coded for by the A subregion of the d, b, p, and q haplotypes (ie. I-Ad,b,p,q).
  Results of flow cytometric analysis
  (Tissue distribution):
  Mouse Strain: BALB/c
  Cell concentration : 1x10e6 cells per test
  Antibody concentration used: 0.1 μg/10e6 cells
  Isotypic control: PE Mouse IgG2a
  Cell source percentage of cells stained above control:
  Spleen 52.0% (see picture below)
  Lymph Node 13.5%
  (Strain distribution ):
  Antibody concentration: 0.2 μg/10e6 cells
  Strains Tested: A.TH, A.TL, C3H/He, C57BL/6, DBA/1
  Positive: C57BL/6, DBA/1
  Negative: A.TH, A.TL, C3H/He
• Formulation: PBS with 0.02% sodium azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE
  State: Liquid Ig raction
  Label: R-Phycoerythrin
• Concentration: 0.1 mg/ml
• Purification: Protein G affinity chromatography
• Conjugation: PE
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  Method:
  1. Prepare a 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 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 0.2 - 0.1 μg of CL090R 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 ofepropidium 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).