T Cell Receptor (TCR) V alpha-2 Rat Monoclonal Antibody [Clone ID: B20.1]

CAT#: AM31862FC-N

T Cell Receptor (TCR) V alpha-2 rat monoclonal antibody, clone B20.1, FITC

Product Images

Figure 1. Cell Source: Mouse Lymph Node. Percentage of cells stained above control: 7.9 %

Specifications

Product Data

• Clone Name: B20.1
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
  This clone has also been reported to work in Immunoprecipitation. (1,2)
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Purified soluble α/β T cell receptor from the cytotoxic T cell clone, KB5-C20. (1)
• Specificity: This antibody reacts with Mouse T-Cell Receptor (TCR) Vα2 chains (1), and recognizes the majority of the TCR Vα2 subfamily in mice carrying the a, b and c haplotypes 1,2.
  It also reacts with the products of T Cell Receptor, Vδ8 due to the high degree of homology (1).
• Formulation: PBS containing 0.09% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: FITC
  State: Liquid purified IgG fraction.
• Concentration: 0.1 mg/ml
• Conjugation: FITC
• Background: The TCR alpha chain complexes with the TCR beta chain to form the T cell receptor in 95% of T cells, whereas the remaining 5% of T cells express gamma and delta chains (γ/δ). TCR Vα2 is a distinct TCR subfamily found in mice having the a, b, and c haplotypes.
• Note: Protocol: Flow Cytometry Analysis:
  
  Method:
  1. Prepare cell suspension in Media A. For cell reparations, deplete the red blood cell population with Lympholyte®-M cell separation medium.
  2. Wash 2 times.
  3. Resuspend the cells to a concentration 2x10e7 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1x10e6 cells, representing one test).
  4. To each tube add ~1.0 µg of this antibody AM31862FC-N per 1x10e6 cells.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate tubes at 4°C for 30-60 minutes. (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 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:
  (Representative Histogram in Figure.1)
  Mouse Strain: C57BL/6
  Cell Concentration : 1x10e6 cells per test
  Antibody Concentration Used: 0.12 µg/10e6 cells
  Isotypic Control: FITC Rat IgG2