Biolegend, 430230, FluoroSpot MAX™ Deluxe Set Human TNF-α/IL-6

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 IL-6 (Interleukin-6), also known as B-cell Stimulatory Factor 2 (BSF2), Interferon beta-2 (IFN-β-2), Hybridoma Growth Factor (HGF), is a cytokine, which is expressed by many different cell types such as macrophages, T-cells, osteoblasts, endothelial cells, and hepatocytes. It is involved in both pro-inflammatory and anti-inflammatory responses. In particular, IL-6 has critical roles in multiple different biological processes including but not limited to B-cell maturation, osteoclast formation, nerve cell differentiation, muscle healing and hypertrophy. Thus, abnormal level of expression and activity of IL-6 can result in a wide range of diseases from chronic inflammation, cancer, to neurodegenerative diseases. BioLegend’s FluoroSpot MAX™ Deluxe Set contains pre-optimized essential reagents and buffers for the simultaneous detection of human TNF-α and IL-6 secreting cells. In this assay, anti-human TNF-α and IL-6 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 IL-6 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 IL-6-secreting cells, enabling precise quantification of both single cytokine-producing cells and dual cytokine-producing cells within the sample.