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

Specifications

Product Data
Clone Name	V65
Applications	FC, IHC
Recommend Dilution	Flow cytometry. Immunohistochemistry on frozen sections.
Reactivity	Rat
Host	Mouse
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.
Isotype	IgG1
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	lot specific
Conjugation	Biotin
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
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%
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.