Human IFN-gamma Alexa Fluor® 750-conjugated Antibody Summary
Accession # AAP20098.1
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of IFN‑ gamma in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with 50 ng/mL PMA, 1 ug/mL Ionomycin, and 3 uM Monensin for overnight were stained with Mouse Anti-Human CD3 epsilon PE‑conjugated Monoclonal Antibody (Catalog # FAB100P) and either (A) Mouse Anti-Human IFN‑ gamma Alexa Fluor® 750‑conjugated Monoclonal Antibody (Catalog # IC2851S) or (B) Mouse IgG2A Alexa Fluor® 750 Isotype Control (Catalog # IC003S). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
- 12 months from date of receipt, 2 to 8 °C as supplied.
Interferon-gamma (IFN-gamma ), also known as type II or immune interferon, exerts a wide range of immunoregulatory activities and is considered to be the prototype proinflammatory cytokine (1, 2). Mature human IFN-gamma exists as a non-covalently linked homodimer of 20-25 kDa variably glycosylated subunits (3). It shares 90% amino acid (aa) sequence identity with rhesus IFN-gamma, 59-64% with bovine, canine, equine, feline, and porcine IFN‑ gamma, and 37-43% with cotton rat, mouse, and rat IFN-gamma. IFN-gamma dimers bind to IFN-gamma RI (alpha subunits) which then interact with IFN-gamma RII (beta subunits) to form the functional receptor complex of two alpha and two beta subunits. Inclusion of IFN-gamma RII increases the binding affinity for ligand and the efficiency of signal transduction (4, 5). IFN-gamma is produced by a variety of immune cells including monocytes, NK cells, gamma δ T cells, CD8+ Tcells, multiple T cell subsets (inlcuding Th1, CCR7- TEM and CD103+CD69+ TRM cells plus proinflammatory FoxP3 regulatory T cells (6-12). It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects (6, 13, 14). In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation (15, 16). The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis (7). Finally, IFN-gamma regulates blood cell production, particularly during immune challenge. In particular, IFN-gamma appears to promote monocyte production while depressing neutrophil, B cell and eosinophil production (17).
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