ELISpot MAX™ Deluxe Set Human TNF-α

Human TNF-α (Tumor Necrosis Factor α), also known as Cachectin, Cytotoxic Factor (CF), is a cytokine which has important roles in regulating a wide range of biological functions from immune activation, cellular differentiation, proliferation, apoptosis, and energy metabolism. It can be expressed and secreted by macrophages, neutrophils, T-cells, NK cells, and a few other cell types. TNF-α can bind to two receptors: TNFR1 and TNFR2. Signaling through TNFR1 is mostly pro-inflammatory, while TNFR2 signaling is mainly anti-inflammatory and proliferative. Because of its diverse functions, dysregulation or TNF-α expression and activity has been implicated in many diseases from autoimmune, cancer, to neurodegeneration. On the other hand, there is also interest in using TNF-α as an immunostimulant drug to enhance the anti-cancer activity of monoclonal antibodies.

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 TNF-α secreting cells. A monoclonal antibody specific for human 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 37oC for a specific period of time. During this period, capture antibody binds to human TNF-α from the secreting cells in the vicinity. Then, cells are removed, and a biotinylated anti-human 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 human TNF-α secreting cells. The end result is visible spots, which corresponds to footprints of TNF-α secreting cells, on the membrane surface.