SignalKine™ Human TNF-α Chemiluminescent Sandwich ELISA Kit from Cell Signaling Technology (CST) is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects human TNF-α (hTNF-α) in multiple matrices. Unknown samples being tested for hTNF-α and hTNF-α standards are added to low volume microwells, where the hTNF-α is captured by the coated hTNF-α Rabbit mAb. Following a washing step, a biotinylated hTNF-α Detection Rabbit mAb is added to detect the captured hTNF-α. HRP-linked Streptavidin is then used for detection of the biotinylated hTNF-α Detection Rabbit mAb. A chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU) is proportional to the quantity of hTNF-α in the sample.
SignalKine™ Human TNF-α Chemiluminescent Sandwich ELISA Kit detects hTNF-α in multiple matrices that can be quantified by generating a standard curve with the recombinant hTNF-α protein standard provided. The hTNF-α standard range is from 2.4 to 10,000 pg/ml. Samples containing higher levels of hTNF-α can be diluted to fit into the standards range.
TNF-α, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein (1,2). Membrane-bound TNF-α is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer (2). Both membrane and soluble forms of TNF-α are biologically active. TNF-α is produced by a variety of immune cells including T cells, B cells, NK cells, and macrophages (1). Cellular response to TNF-α is mediated through interaction with receptors TNF-R1 and TNF-R2 and results in activation of pathways that favor both cell survival and apoptosis depending on the cell type and biological context. Activation of kinase pathways (including JNK, Erk1/2, p38 MAPK, and NF-κB) promotes the survival of cells, while TNF-α-mediated activation of caspase-8 leads to programmed cell death (1,2). TNF-α plays a key regulatory role in inflammation and host defense against bacterial infection, notably Mycobacterium tuberculosis (3).
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