TCR alpha/beta antibody (Biotin)

Catalog No. ABIN114324

Product Details anti-TCR alpha/beta Antibody (Biotin)

Target: TCR alpha/beta (T Cell Receptor alpha/beta (TCR alpha/beta))

Reactivity: Mouse

Host: Hamster

Clonality: Monoclonal

Conjugate: This TCR alpha/beta antibody is conjugated to Biotin

Application: Flow Cytometry (FACS)

Purification: Protein G Chromatography

Immunogen: Affinity Purified D0-11.10 TCR Donor: Armenian Hamster Fusion Partner: Murine myeloma variant P3X63Ag.653

Clone: H57-597

Isotype: IgG

Application Details

Application Notes: Flow cytometry. Immunoprecipitation. This clone has also been reported to work with frozen sections and Western Blotting.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation 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 thissuspension 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. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1: 500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protectedfrom 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 propidiumiodide 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 (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: BALB/cCell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 1. 0 µg/10e6 cellsIsotypic Control: Biotin Hamster IgGCell Source Percentage of cells stained above control: Thymus: 62. 6% see FIGURE 1Splenic T Cells: 76. 2%Strain Distribution by Flow Cytometry Analysis: Cell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 1. 0 µg/106 cellsStrains Tested: BALB/c, C57BL/6, CBA/J, C3H/He, AKRPositive: BALB/c, C57BL/6, CBA/J, C3H/He, AKRNegative: none

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: PBS,0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for TCR alpha/beta

Target: TCR alpha/beta (T Cell Receptor alpha/beta (TCR alpha/beta))

Alternative Name: T Cell Receptor / TCR alpha/beta (TCR alpha/beta Products)

Background: Synonyms: T-Cell Receptor alpha, T-Cell Receptor alpha beta, T-Cell Receptor beta, TCRA, TCRB