MHC Class I H-2Ld Mouse Monoclonal Antibody [Clone ID: 30-5-7S]

CAT#: CL060F

MHC Class I H-2Ld mouse monoclonal antibody, clone 30-5-7S, FITC

Product Images

Figure 1. Balb/c Lymph Node labelled with CL060F including Isotype Control (Shaded).

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  Figure 2.

Specifications

Product Data

• Clone Name: 30-5-7S
• Applications: Assay, CT, FC
• Recommended Dilution: Flow Cytometry (see protocol).
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Recipient: BALB/c-H-2dm2
  Donor: BALB/c Spleen cells
  Fusion Partner: SP2/0.Ag14
• Specificity: This monoclonal antibody detects the public specificity H-2.65 of the H-2Ld antigen.
  This antibody also recognizes H-2Dq and H-2Lq molecules.
• Formulation: PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as stabilizing protein to bring the protein concentration to 4-5 mg/ml
  Label: FITC
  State: Liquid purified IgG fraction
• Conjugation: FITC
• Synonyms: H2-L
• 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 2.0 µg of this Antibody CL060F 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 in Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration: 1x10e6 cells/test
  Antibody Concentration Used: 2.0 µg/10e6 cells
  Isotype Control: FITC Mouse IgG2a
  
  Cell Source - % of cells stained above Control:
  Spleen: 92.4%
  Thymus: 54.4%
  Lymph Node: 95.3%
  
  Strain Distribution:
  Procedure: As above
  Antibody Concentration: 1µg/10e6 cells
  Strains Tested: See Figure 2