K4779-100? TNF-alpha Microplate (Item A) coated with anti-human TNF-alpha, 96 wells ? Wash Buffer Concentrate (20x) (Item B) ? Standards (Item C) (recombinant human TNF-alpha) ? Assay Diluent A (Item D) for Standard/Sample (serum/plasma) diluent (0.09% sodium azide as preservative) ? Assay Diluent B (Item E) for Standard/Sample (cell culture medium/urine) diluent. (5x concentrated) ? Detection Antibody TNF-alpha (Item F), biotinylated anti-human TNF-alpha (each vial enough for half plate) ? HRP-Streptavidin Concentrate (Item G), 600x concentrated ? TMB One-Step Substrate Reagent (Item H) 3,3’,5,5’-tetramethylbenzidine (TMB) in buffer solution ? Stop Solution (Item I), 0.2 M sulfuric acid ? Serum & plasma (human TNF-alpha concentration is pretty low in normal serum & plasma, it may not be detected in this assay) ? Cell culture supernatants ? Urine TNF-alpha (human) ELISA Kit: Colorimetric Assay for Quantitative measurement of human TNF-alpha in Serum, Plasma, urine & Cell culture supernatants. Detection Range: 30 pg/ml - 6000 pg/ml. 100 assays. BioVision’s Human TNF-alpha (Tumor necrosis factor-alpha) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human TNF-alpha. This assay employs an antibody specific for human TNF-alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and TNF-alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human TNF-alpha antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF-alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. This ELISA kit shows no cross-reactivity with any of the cytokines tested e.g., human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO- alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1alpha, IL-1beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1alpha, MIP-1beta, NAP-2, PDGF, PF-4, PARC, SCF, SDF-1α, TIMP-1, TIMP-2, TNFβ, TGFβ1, TGFβ2, TGFβ3, VEGF. The minimum detectable dose of TNF-alpha is typically less than 30 pg/ml. Detection Range: 30 pg/ml – 6,000 pg/ml. The intra-Assay reproducibility is CV<10% & inter-Assay is CV<12%.