Biolegend, 430228, FluoroSpot MAX™ Deluxe Set Human TNF-α/IFN-γ

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. TNF-α is expressed and secreted by macrophages, neutrophils, T-cells, NK cells, and various 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. 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 multifunctional nature, dysregulation of IFN-γ expression and activity has been implicated in many diseases such as autoimmune and cancer. BioLegend’s FluoroSpot MAX™ Deluxe Set contains pre-optimized essential reagents and buffers for the simultaneous detection of human TNF-α and IFN-γ secreting cells. In this assay, anti-human TNF-α and IFN-γ capture antibodies are coated on a clear-bottom microplate. Cells (stimulated or unstimulated) are added to the wells and incubated, allowing secreted TNF-α and/or IFN-γ to bind to nearby capture antibodies. After incubation, cells are removed, and fluorescent conjugated detection antibodies are added. The resulting spots represent the "footprints" of individual TNF-α and IFN-γ-secreting cells, enabling precise quantification of both single cytokine-producing cells and dual cytokine-producing cells within the sample.