Human TNF alpha ELISPOT Kit is a ELISPOT Kit for the sensitive detection of Human TNF alpha production in a single cell suspension.
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISPOT | Reactivity Reacts | Dilution info - | Notes - |
The TNF protein, primarily secreted by macrophages, binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It induces cell death in specific tumor cell lines and acts as a potent pyrogen, causing fever directly or by stimulating interleukin-1 secretion. TNF is implicated in cachexia induction and can stimulate cell proliferation and differentiation under certain conditions. It impairs regulatory T-cells (Treg) function in rheumatoid arthritis patients through FOXP3 dephosphorylation, upregulating protein phosphatase 1 (PP1), which dephosphorylates 'Ser-418' of FOXP3, inactivating FOXP3 and leading to defective Treg cells. TNF is a key mediator of cell death in the anticancer effect of BCG-stimulated neutrophils with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line. It induces insulin resistance in adipocytes by inhibiting insulin-induced IRS1 tyrosine phosphorylation and glucose uptake. TNF plays a role in angiogenesis by inducing VEGF production with IL1B and IL6. Additionally, the TNF intracellular domain (ICD) form stimulates IL12 production in dendritic cells. This supplementary information is collated from multiple sources and compiled automatically.
TNFA, TNFSF2, TNF, Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a
Human TNF alpha ELISPOT Kit is a ELISPOT Kit for the sensitive detection of Human TNF alpha production in a single cell suspension.
The ELISPOT assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.
Abcam Elispot assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.
Principal:
After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.
Recognizes natural human TNF alpha
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