Biolegend, 430924, ELISpot MAX™ Deluxe Set Mouse TNF-α

TNF-α (Tumor Necrosis Factor α), also known as Cachectin, is a cytokine belonging to the Tumor Necrosis Factor superfamily. It is produced by various cell types, including macrophages, monocytes, neutrophils, T cells, NK cells, and epithelial cells. TNF-α can bind to two receptors: TNFR1 and TNFR2. TNF-α signaling plays a central role in the inflammatory response, lymphoid cell development, and lipid metabolism. Additionally, TNF-α is a pathological component in numerous autoimmune diseases, including but not limited to Crohn’s disease, asthma, and rheumatoid arthritis. It was also reported to be involved in some cancers and obesity. 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 mouse TNF-α secreting cells. An antibody specific for mouse TNF-α 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 mouse TNF-α from the secreting cells in the vicinity. Then, cells are removed, and a biotinylated anti-mouse TNF-α 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 mouse TNF-α secreting cells. The end result is visible spots, which correspond to footprints of TNF-α secreting cells, on the membrane surface.