T Cell Receptor (TCR) gamma/delta Mouse Monoclonal Antibody [Clone ID: V65]

CAT#: SM322B

T Cell Receptor (TCR) gamma/delta mouse monoclonal antibody, clone V65, Biotin

Product Images

LFL2 - Cell Source: Splenic T Cells - Percentage of cells stained above control: 5.3%

Specifications

Product Data

• Clone Name: V65
• Applications: FC, IHC
• Recommended Dilution: Flow cytometry.
  Immunohistochemistry on frozen sections.
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Rat T Blasts.
  Donor: BALB/c spleen.
  Fusion Partner: X63-Ag 8.653.
• Specificity: This monoclonal antibody detects a T cell-specific heterodimeric 48 and 50 kD cell surface protein that is expressed on greater than 90% of CD3+ abTCR- rat peripheral T lymphocytes, and identifies a dense network of dendritic cells in the epidermis as g/d T cells. Immobilized this antibody induces a strong proliferative response in gd T cell cultures supplemented with either IL-2 or IL-4.
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid purified Ig
• Concentration: 0.1 mg/ml
• Conjugation: Biotin
• Synonyms: TCRG, TCRD, T-Cell Receptor gamma, T-Cell Receptor delta, T-Cell Receptor gamma delta
• 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®-Rat 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 0.1-0.2 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-PE) at a 1:20 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:
  Rat Strain: Fischer
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.2 µg/10e6 cells
  Isotypic Control: Biotin Mouse IgG1
  
  Cell Source Percentage of cells stained above control:
  Thymus: 3.6%
  Splenic T Cells: 5.3%