T Cell Receptor (TCR) alpha/beta Hamster Monoclonal Antibody [Clone ID: H57-597]

CAT#: CL075F

T Cell Receptor (TCR) alpha/beta hamster monoclonal antibody, clone H57-597, FITC

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Specifications

Product Data

• Clone Name: H57-597
• Applications: FC
• Recommended Dilution: Flow Cytometry.
  Immunohistochemistry on Frozen Sections: Clone has reportedly worked with this application (See also Reference 6).
• Reactivities: Mouse
• Host: Hamster
• Isotype: IgG
• Clonality: Monoclonal
• Immunogen: Affinity-purified DO-11.10 TCR from Armenian Hamster.
  Fusion Partner: Mouse myeloma variant P3X63 Ag.653.
• Specificity: This anti-Mouse antibody T cell receptor monoclonal antibody reacts with the surface of all ab TCR bearing cells and does not react with receptors on gd TCR positive T cells. This monoclonal antibody when used in an immobilized form was able to activate all ab TCR bearing T cell hybridomas tested to produce IL-2.
  Use of this antibody in conjunction with an anti-CD3e monoclonal antibody allows for accurate measurements of the mutually exclusive sub-populations of ab TCR and gd TCR bearing T cells.
• Formulation: PBS, 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: FITC
  State: Liquid purified IgG fractiom
• Concentration: lot specific
• Purification: Protein G Chromatography
• Conjugation: FITC
• Storage: Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
  This product is photosensitive and should protected from light.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Synonyms: TCRA, TCRB, T-Cell Receptor alpha, T-Cell Receptor beta, T-Cell Receptor alpha beta
• Note: Protocol: Floy 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 2x10⁷ cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10⁶ cells, representing 1 test).
  4. To each tube, add 0.2-0.1 µg* of this Ab per 10⁶ 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: 1x10⁶ cells per test
  Antibody Concentration Used: 0.2 µg/10⁶ cells.
  Isotypic Control: FITC Hamster IgG
  
  Cell Source: Percentage of Cells Stained Above Control
  Thymus: 76.9% see Figure A
  Splenic T Cells: 90.3% see Figure B
  
  Strain Distribution by Flow Cytometry Analysis:
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 µg/10⁶ cells
  Strains Tested: C57BL/6, CBA/J, AKR, BALB/c, C3H/He
  Positive: C57BL/6, CBA/J, AKR, BALB/c, C3H/He
  Negative: none