Biolegend, 430124, ELISpot MAX™ Deluxe Set Human IFN-γ

Human IFN-γ (Interferon-gamma) is a multifunctional cytokine which is the sole member of the class II Interferon. Although a variety of immune cells can express IFN-γ, most of IFN-γ is produced by T-cells and NK cells. It has critical roles in many processes including, but not limited to, anti-viral response, regulating Th1/Th2 balance, enhancing antigen presentation, and controlling cellular proliferation and apoptosis. IFN-γ can bind to 2 receptors: IFNGR1 and IFNGR2, which transduce the signal downstream through the JAK-STAT pathway. Because of its multifunction nature, dysregulation of IFN-γ expression and activity has been implicated in many diseases such as autoimmune and cancer. BioLegend’s ELISpot MAX™ Deluxe Set contains pre-optimized essential reagents and additional buffers and solution for Enzyme-Linked Immunosorbent Spot assay. This kit is designed for detection of human IFN-γ secreting cells. A monoclonal antibody specific for human IFN-γ is coated on ethanol-activated 96-well PVDF membrane plate as the capture. Stimulated and/or unstimulated cells are pipetted directly into the wells, and then incubated at 37 ° C for a specific period of time. During this period, capture antibody binds to human IFN-γ from the secreting cells in the vicinity. Then, cells are removed, and a biotinylated anti-human IFN-γ detection antibody is added, followed by Avidin-HRP. Subsequently, TMB membrane substrate is added. The TMB membrane substrate reacts with HRP to form a blue-black precipitate at the sites of human IFN-γ secreting cells. The end result is visible spots, which correspond to footprints of IFN-γ secreting cells on the membrane surface.