Monoclonal Antibody to T Cell Receptor (TCR

BM2467 OriGene Technologies Inc. OriGene EU

Acris Antibodies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-858-888-7900 Phone: +49-5221-34606-0 Fax: +1-858-888-7904 Fax: +49-5221-34606-11 [email protected] [email protected]

Monoclonal Antibody to T Cell Receptor (TCR) gamma/delta - Purified Alternate names: T-Cell Receptor delta, T-Cell Receptor gamma, T-Cell Receptor gamma delta, TCRD, TCRG Catalog No.: BM2467 Quantity: 0.1 mg Concentration: 0.2 mg/ml (prior to lyophilization) Background: T cell receptors (TCR) recognize foreign antigens which have been processed as small peptides and bound to major histocompatibility complex (MHC) molecules at the surface of antigen presenting cells (APC). Each T cell receptor is a dimer consisting of one alpha and one beta chain or one delta and one gamma chain. This region represents the germline organization of the T cell receptor beta locus. The beta locus includes V (variable), J (joining), diversity (D), and C (constant) segments. During T cell development, the beta chain is synthesized by a recombination event at the DNA level joining a D segment with a J segment; a V segment is then joined to the D-J gene. The C segment is later joined by splicing at the RNA level. The TCR gamma/delta heterodimer of the CD3/T-cell receptor complex is clonotypic and consists of Ig-like variable and constant domains. It is expressed during thymopoiesis and on the small subpopulation of gamma/delta TCR expressing T lymphocytes in the periphery. The majority of T cells present in some epithelial tissues are gamma/delta+ and have limited receptor diversity. The TCR gamma/delta heterodimer recognizes peptide antigen bound to MHC antigens, and subsequent signal transduction mediated by the invariant chains leads to T cell activation. Host / Isotype: Mouse / IgG1 Recommended SM10P (for use in human samples), AM03095PU-N Isotype Controls: Clone: Immu510 Immunogen: Soluble gamma/delta T-cell receptor. Format: State: Lyophilized purified Ig fraction Purification: Affinity Chromatography on Protein A Buffer System: PBS with 1 mg/ml BSA as stabilizer without preservatives Reconstitution: Restore with 0.5 ml distilled water. Applications: Flow Cytometry: 2 µg/5x10e5 cells/test or 10 µl of the reconstituted purified antibody. As this antibody recognizes a rare population, around 3.6% of peripheral blood lymphocytes (4), it is often preferable to use double staining experiments with another T- cell marker (CD2, CD3, CD4, CD8). For protocol see below. Western blot: Recognizes the gamma/delta complex under non-reducing conditions and the isolated delta chain on reducing conditions.

For research and in vitro use only. Not for diagnostic or therapeutic work. OG/20130615 Material Safety Datasheets are available at www.acris-antibodies.com or on request. Acris Antibodies is now part of the OriGene family. Learn more at www.origene.com 1 / 3 BM2467: Monoclonal Antibody to T Cell Receptor (TCR) gamma/delta - Purified Immunoprecipitation: Precipitates the three isoforms of the gamma/delta T-cell receptor (2). Immunofluorescence: Gives excellent results on frozen Sections (4). Immunohistochemisty on Frozen Sections. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user. Specificity: This clone recognizes all Human gamma/delta T cells regardless the variable genes or junction regions they express as assessed by Flow Immunofluorescence studies on polyclonal gamma/delta T-cell lines as well as gamma/delta T-cell clones (1-5). It has been characterized in reference 1. It recognizes 1.6 to 8.9% of peripheral CD3 positive lymphocytes from 7 healthy adult donors. Species: Human. Other species not tested. Storage: Store lyophilized at 2-8°C. After reconstitution store (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: one year from despatch. General Readings: 1. Davodeau, F., et al., "Secretion of disulfide-linked human T-cell receptor gamma/delta heterodimers", 1993, J. Biol. Chem. 268, 15455-15480. 2. Davodeau F, Peyrat MA, Houde I, Hallet MM, De Libero G, Vié H, et al. Surface expression of two distinct functional antigen receptors on human gamma delta T cells. Science. 1993 Jun 18;260(5115):1800-2. PubMed PMID: 8390096. 3. Davodeau F, Peyrat MA, Romagné F, Necker A, Hallet MM, Vié H, et al. Dual T cell receptor beta chain expression on human T lymphocytes. J Exp Med. 1995 Apr 1;181(4):1391-8. PubMed PMID: 7699325. 4. Peyrat MA, Davodeau F, Houde I, Romagné F, Necker A, Leget C, et al. Repertoire analysis of human peripheral blood lymphocytes using a human V delta 3 region-specific monoclonal antibody. Characterization of dual T cell receptor (TCR) delta-chain expressors and alpha beta T cells expressing V delta 3J alpha C alpha-encoded TCR chains. J Immunol. 1995 Sep 15;155(6):3060-7. PubMed PMID: 7673722. 5. Thibault G, Bardos P. Compared TCR and CD3 epsilon expression on alpha beta and gamma delta T cells. Evidence for the association of two TCR heterodimers with three CD3 epsilon chains in the TCR/CD3 complex. J Immunol. 1995 Apr 15;154(8):3814-20. PubMed PMID: 7706721. Protocols: Double staining is also possible with purified unlabeled antibody form using the following protocol:

Double labeling protocol using lyophilized unconjugated form with a CD3 (Phyco conjugated).

1. To 100 µl of whole blood, add 10ul of the reconst. purified antibody. Incubate 15 minutes at room temperature (18-25°C). 2. Add 3ml of PBS/BSA/NaN3. Centrifuge 5 minutes at 1200 rpm, discard supernatant. 3. Add 100 µl of secondary antibody F(ab')2 goat anti-mouse Ig conjugated to FITC at usual dilution in PBS/BSA/NaN3. Incubate 15 minutes at room temperature. 4. Repeat step 2 (washing). 5. Resuspend cells in 100 µl of PBS/BSA/NaN3 containing 1 mg/ml of total mouse Ig (to saturate eventual free sites of the goat anti-mouse FITC). Incubate 5 minutes at room temperature. 6. Without washing, add 20 µl of the CD3 PE. Incubate 15 minutes at room temperature. 7. Repeat step 2 (washing).