Swine TNF-α ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for TNF-α has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any TNF-α present is bound by the immobilized antibody and the detection antibody specific for TNF-α is binds to the combination of capture antibody-TNF-α in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of TNF-α present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven TNF-α standard dilutions and TNF-α sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.

Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.

GENE ID	397086
SWISS PROT	P23563
SYNONYMS	TNFa

Materials Supplied

Kit Components	96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with antibody to Swine TNF-α (8-12)	1 plate
Swine TNF-α standard lyophilized, 2000pg/ml upon reconstitution	2 vials
Concentrated Biotin-Conjugate anti-Swine TNF-α antibody	2 vials
Streptavidin-HRP solution	2 vials
Standard /sample Diluent	1 bottle
Biotin-Conjugate antibody Diluent	1 bottle
Streptavidin-HRP Diluent	1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20)	1 bottle
Substrate Solution	1 vial
Stop Solution	1 vial
Adhesive Films	4 pieces
Product data sheet	1 copy

Storage

Store at 2 - 8°C

Performance Characteristics

REPEATABILITY	The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY	The minimum detectable dose was 15pg/mL.
ASSAY RANGE	31.25 - 2000 pg/mL
SPECIFICITY	This assay recognizes both natural and recombinant Swine TNF-α. The factors listed below were prepared at 70ng/ml in standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.

Factors assayed for cross-reactivity

Recombinant human	TNF-β, GM-CSF, s TNF RI, s TNF RII, s TNF RII/Fc
Recombinant mouse	TNF-α（truncated, s TNF RI, s TNF RII IL-4

Data Analysis Assistance

We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKW1007.

https://www.myassays.com/bioss-swine-tnf-%ce%b1-elisa-kit.assay