Description
The 536 monoclonal antibody specifically recognizes Vγ 3† T-cell Receptor (TCR)-bearing T lymphocytes, which are the predominant γδ TCR-bearing cells in the early fetal thymus and the adult epidermis of euthymic mice. The first T cells to mature in the embryonic thymus express the Vγ 3 and Vδ 1 TCR chains, and their development is dependent upon IL-7. There is evidence that the Vγ 3 TCR-bearing fetal thymocytes are the precursors of the majority of dendritic epidermal T cells (DEC), which may be replenished in the adult by proliferation in situ rather than by seeding from primary lymphoid organs. Although the Vγ 3 TCR is almost exclusively found in the DEC population, it has been shown that the homing of DEC to the epidermis does not require expression of the Vγ 3 gene segment. Vγ 3 TCR-bearing dermal dendritic cells have also been described. Vγ 3 TCR has also been found on a subset of T lymphocytes in the lactating mammary gland and at the site of antigenic challenge in contact-sensitized mice. Plate-bound 536 antibody activates Vγ 3 TCR-bearing T cells, and Fab fragments of mAb 536 block in vitro stimulation of DEC by keratinocytes. † Please note that the Vγ 3 designation correlates with the nomenclature of Garman, Doherty, and Raulet; the Vγ 5 designation of Heilig and Tonegawa is equivalent.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349). When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values. For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BB700 under optimal conditions that minimize unconjugated dye and antibody.
1. This antibody was developed for use in flow cytometry.
2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
3. Researchers should determine the optimal concentration of this reagent for their individual applications.
4. An isotype control should be used at the same concentration as the antibody of interest.
5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
9. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
10. Cy is a trademark of GE Healthcare.
