T Cell Receptor (TCR) gamma/delta Hamster Monoclonal Antibody [Clone ID: GL3]

CAT#: SM093C

T Cell Receptor (TCR) gamma/delta hamster monoclonal antibody, clone GL3, PE-Cy5

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Specifications

Product Data

• Clone Name: GL3
• Applications: FC
• Recommended Dilution: Flow cytometry.
• Reactivities: Mouse
• Host: Hamster
• Isotype: IgG
• Clonality: Monoclonal
• Specificity: This monoclonal antibody reacts with the surface on all gamma delta TCR bearing cells and does not react with receptors on alpha beta TCR positive cells. It is thought that this clone may be specific for a determinant present on Cδ 7. The gammadelta T cell receptors are present on murine CD4-CD8- thymocytes, peripheral T cells, intestinal CD8+ intraepithelial lymphocytes and Thy 1+ dendritic epidermal cells in the skin.
  Use of this antibody in conjunction with an anti-CD3 monoclonal antibody (anti-CD3epsilon Monoclonal Antibody ) allows for accurate measurements of the mutually exclusive sub-populations of gamma delta TCR and alpha beta TCR bearing T cells. This anti mouse gamma delta TCR monoclonal antibody has also been used successfully for the characterization of murine intraepithelial lymphocytes.
• Formulation: PBS, 0.09% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE-Cy5
  State: Liquid purified Ig
  Absorption emission: 488 nm/ 667 nm
• Concentration: 0.2 mg/ml
• Conjugation: PE-Cy5
• Synonyms: TCRG, TCRD, T-Cell Receptor gamma, T-Cell Receptor delta, T-Cell Receptor gamma delta
• Note: PE-Cy5 conjugates require a 650 nm long pass filter in the FL3 channel. FL2-FL3 compensation will be in the range of 1%.
  
  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 ~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.
  (It is recommended that the tubes are protected from light, since most flurochromes 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:
  (Representative Histogram)
  Mouse Strain: BALB/c
  Cell Concentration: 1 x 10e6 cells per test
  Antibody Concentration used: 10µl /10e6
  Isotypic Control: Hamster IgG Pe-Cy5