MHC Class I H2 Kd/Dd Mouse Monoclonal Antibody [Clone ID: 34-1-2S]

CAT#: AM08081FC-S

MHC Class I H2 Kd/Dd mouse monoclonal antibody, clone 34-1-2S, FITC

Product Images

Cell Source: Spleen Percentage of cells stained above control: 95.1%

• 

Specifications

Product Data

• Clone Name: 34-1-2S
• Applications: FC
• Recommended Dilution: Flow Cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: BDF splenocytes
  Donor: C3H spleen
  Fusion Partner: Sp2/0-Ag14
• Specificity: Anti H-2KdDd mAb reacts with both H-2Kd and H-2Dd products. The antibody also cross reacts with Kbsrpq. This K-D cross reaction indicates the presence of shared specificities between the two separate H-2 regions.
• Formulation: PBS containing 0.02% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein
  Label: FITC
  State: Liquid purified Ig fraction
  Label: Fluorescein isothiocyanate isomer 1
• Concentration: lot specific
• Purification: Affinity chromatography on Protein G
• Conjugation: FITC
• Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• 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 0.5 - 0.2 μg of this 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. (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 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: BALB/c
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.5 μg/10e6 cells
  Isotypic Control: FITC Mouse IgG2a
  Percentage of cells stained above control:
  Thymus 91.9%
  Spleen 95.1%
  Lymph Node 100%