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

CAT#: CL053F

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

Product Images

FIGURE1: LFL1 - Cell Source: Spleen - Percentage of Cells Stained Above Control: 97.9%

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  FIGURE2

Specifications

Product Data

• Clone Name: 34-5-8S
• Applications: Assay, FC
• Recommended Dilution: Flow cytometry.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Recipient: C3H/HeJ
  Immunocyte Donor: B6 x DBA/2 spleen cells
  Fusion Partner: SP2/0.Ag14
• Specificity: This anti-mouse H-2Dd monoclonal antibody 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 the H-2Dd(H-2.4) antigen from the appropriate strains of mice.
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: FITC
  State: Liquid purified Ig
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: FITC
• 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 cell suspension in Media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation media.
  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 mg 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. (It is recommended that 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 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).
  
  Results - Tissue Distribution by Flow Cytometry Analysis:
  Donor: BALB/c
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration: 1 µg/10e6 cells
  Isotypic Control: FITC Mouse IgG2a, k
  
  STRAIN DISTRIBUTION:
  Procedure: As above
  Antibody Concentration: 1µg/10e6 cells
  Strains Tested: see Figure 2