TCR alpha/beta Monoclonal Antibody (IP26), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

• Description: The IP26 monoclonal antibody reacts with the alpha beta chain of human TCR.The alpha beta TCR is expressed by the majority of peripheral T cells
• Applications Reported: This IP26 antibody has been reported for use in flow cytometric analysis
• Applications Tested: This IP26 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells
• This can be used at 5 μL (0.5 μg) per test
• A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5
• We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative
• Please make sure that your instrument is capable of detecting this fluorochrome
• Fixation: Samples can be stored in IC Fixation Buffer (cat
• 00-8222) (100 μL cell sample + 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat
• 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically
• The T cell antigen receptor (TCR) consists of a ligand-specific alpha/beta heterodimer non-covalently associated with five invariant chains including the CD3 gamma/delta/eta and zeta subunits, all of which are required for efficient surface expression
• T cell activation through the TCR induces cellular differentiation and/or proliferation and the production of lymphokines and cytokines
• Both the CD3 and TCR zeta subunits are proposed to be responsible for the intracellular signal transduction events
• Majority of T cells present in the blood, lymph and secondary lymphoid organs express TCR alpha/beta heterodimers, whereas the T cells expressing TCR gamma/delta heterodimers are localized mainly in epithelial tissues and at the sites of infection
• The subunits of TCR heterodimers are covalently bonded and associate with the CD3 subunits in the endoplasmic reticulum to form functional TCR-CD3 complex
• Lack of expression of any of the chains is sufficient to stop cell surface expression
• The ability of T cell receptors (TCR) to discriminate foreign from self-peptides presented by major histocompatibility complex (MHC) class II molecules is essential for an effective adaptive immune response
• TCR recognition of self-peptides has been linked to autoimmune disease
• Mutant self-peptides have been associated with tumors
• Engagement of TCRs by a family of bacterial toxins know as superantigens has been responsible for toxic shock syndrome
• Autoantibodies to V beta segments of T cell receptors have been isolated from patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE)
• The autoantibodies block TH1-mediated inflammatory auto-destructive reactions and are believed to be a method by which the immune system compensates for disease (ref5)
• T Cell and TCR Diversity Most human T cells express the TCR alpha-beta and either CD4 or CD8 molecule (single positive, SP)
• A small number of T cells lack both CD4 and CD8 (double negative, DN)
• Increased percentages of alpha-beta DN T cells have been identified in some autoimmune and immunodeficiency disorders
• Gamma-delta T cells are primarily found within the epithelium
• They show less TCR diversity and recognize antigens differently than alpha-beta T cells
• Subsets of gamma-delta T cells have shown antitumor and immunoregulatory activity.