Human Tumor Necrosis Factor alpha ELISA Kit (A33004)

$715
Freight Charges
Lead Time
5-8 Business Days
Telephone
+1 (314) 370-6046
Mon - Fri, 8am - 4pm AST
Email
orders@antibodies.com
Name
Human Tumor Necrosis Factor alpha ELISA Kit
Assay Type
Sandwich (quantitative)
Principle of Assay
This TNF-α enzyme linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to TNF-α. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated monoclonal antibody preparation specific for TNF-α and incubated. TNF-α, if present, will bind and become immobilized by the antibody pre-coated on the wells and then be “sandwiched” by biotin conjugate. The microtiter plate wells are thoroughly washed to remove unbound TNF-α and other components of the sample. In order to quantify the amount of TNF-α present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Avidin is a tetramer containing four identical subunits that each has a high affinity-binding site for biotin. The wells are thoroughly washed to remove all unbound HRP-conjugated Avidin and a TMB (3,3'5,5' tetramethyl-enzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain TNF-α, biotin conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.

In order to measure the concentration of TNF-α in the samples, this kit includes two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing.) According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D) versus TNF-α concentration (pg/mL). The concentration of TNF-α in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Platform
Microplate
Detection Type
Colorimetric
Sample Type
Serum, plasma, cell culture supernatant, and other biological fluids.
Sensitivity
< 4 pg/ml
Product Range
4-2000 pg/ml
Assay Time
2h 30m - 3h
Specificity
This sandwich ELISA recognises both natural and recombinant human TNF-α.
Reactivity
Human
Cross Reactivity
This assay has shown no cross-reactivity with other cytokines tested.
Recovery
Storage
Store at 2-8°C.
Synonyms
APC1, APC1 protein, Cachectin, DIF, Differentiation inducing factor, Macrophage cytotoxic factor, soluble form, Tnf, TNF a, TNF alpha, TNF macrophage derived, TNF monocyte derived, TNF superfamily member 2 , TNF superfamily, member 2, TNF, macrophage derived, TNF, monocyte derived, TNF-a, TNF-alpha, TNFA, TNFA_HUMAN, TNFSF2, Tumor necrosis factor, Tumor necrosis factor (TNF superfamily member 2) , Tumor necrosis factor ligand superfamily member 2, Tumor Necrosis Factor, Membrane Form, Tumor necrosis factor, soluble form
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
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