LOC102641613 Mouse Monoclonal Antibody [Clone ID: 34-5-8S]

CAT#: CL053B

LOC102641613 mouse monoclonal antibody, clone 34-5-8S, Biotin

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Specifications

Product Data

• Clone Name: 34-5-8S
• Applications: FC
• Recommended Dilution: Flow cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: B6xDBA/2 spleen cells
  Donor: C3H/HeJ spleen
  Fusion Partner: SP2/0.Ag14
• Specificity: This mAb is specific for cells expressing the H-2D antigen coded for by the d haplotype. The reaction pattern of this antibody with a panel of inbred and recombinant haplotypes demonstrates that the antibody detects a private determinant (H-2.4) of the H-2Dd antigen. This antibody can be used to quantitate cells bearing H-2Dd (H-2.4) antigen from the appropriate strains of mice.
• Formulation: PBS , 0.02% NaN3 and EIA grade BSA as a stabalizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: Biotin
  State: Liquid purified Ig
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: Biotin
• Gene Name: H-2 class I histocompatibility antigen, D-D alpha chain
• Database Link:
  Entrez Gene 102641613 Mouse
• Synonyms: H2-D1, H-2D(D)
• 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 0.5-1.0 µg* of this Ab 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 a 1:500 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).
  
  Results - Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration : 1x10e6 cells per tests
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Isotypic Control: Biotin Mouse IgG2a
  
  Cell Source Percentage of cells stained above control:
  Thymus: 40.7 %
  Spleen: 98.3%
  
  Strain Distribution by Flow Cytometry Analysis:
  Cell Concentration : 1x10e6 cells per tests
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Strains Tested: BALB/c, C57BL/6, CBA/J, C3H/He
  Positive: BALB/c
  Negative: C57BL/6, CBA/J, C3H/He