|The TNF-a tnf (Catalog #MBS590025) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase.
The TNF-a tnf product has the following accession number(s) (GI #37210) (NCBI Accession #CAA26669.1) (Uniprot Accession #P01375). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful.
To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
(Or you can also download the PDF Manual
for complete product instructions).
Please refer to the product datasheet for known applications of a given elisa kit. We\'ve tested the Tumor Necrosis Factor alpha (TNF-a) with the following immunoassay(s):
Typical Testing Data/Standard Curve (for reference only)
Principle of the assay: This TNF-alpha 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-alpha. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated monoclonal antibody preparation specific for TNF-alpha and incubated. TNF-alpha, 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-alpha and other components of the sample. In order to quantify the amount of TNF-alpha 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-benzidine) 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-alpha, 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-alpha 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-alpha concentration (pg/mL). The concentration of TNF-alpha in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Samples: Cell Culture Supernatant, serum, plasma, and other biological fluids
Assay Type: Sandwich
Sensitivity: The minimum detectable dose of TNF-? using a standard curve generated with Calibrator Diluent I or Calibrator Diluent II is 2.4pg/mL.