TNF alpha ELISpot Assay Kit (Human)

ELISA Kit

Wet Ice

TNF

233

This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine.

NP_000585.2

Assay Methodology: Quantitative Sandwich ELISpot Immunoassay

Aviva's Human TNF alpha ELISpot Kit is based on standard sandwich ELISA methodology. Capture antibodies highly specific for the analyte of interest are coated to the wells of a PVDF bottomed 96 well microtitre plate either during kit manufacture or in the laboratory. The plate is then blocked to minimise any non-antibody dependent unspecific binding and finally washed before adding the cells to be investigated. Cell suspension and stimulant are added to the coated and blocked microtitre plate and the plate incubated allowing the specific antibodies to bind any analytes produced. Biotinylated detection antibodies are then added which bind to the previously captured analyte. Enzyme conjugated streptavidin is added binding to the detection antibodies. Any excess unbound analyte and antibodies are removed by careful washing. Colour substrate is then applied to the wells resulting in coloured spots which can be quantified using appropriate analysis software or manually using microscopes.