Human TNF-α ELISA Kit
Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
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. Standards and samples are pipetted into the wells and any TNF-α present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for TNF-α is added to the wells and 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
The prototype ligand of the TNF superfamily, TNF-α/TNFSF1A, is a pleiotropic cytokine that plays a central role in inflammation and apoptosis. Human cells known to express TNF-α include B cells, colonic columnar epithelial cells, NK and CD3+CD56+ hepatic natural T cells, macrophages, monocytes and monocyte-derived dendritic cells, CD4+ and CD8+ T cells, mast cells, neutrophils, keratinocytes, plasma cells, and adipocytes.
It is synthesized as a 26 kDa, type II transmembrane protein that is 233 amino acids (aa) in length. It contains a 30 aa cytoplasmic domain, a 26 amino acids transmembrane segment, and a 177 amino acids extracellular region. TNF-α is assembled intracellularly to form a transmembrane, non-covalently-linked homotrimeric protein. The 157 aa residue soluble form of TNF-α (sTNF-α is released from the C-terminus of the transmembrane protein through the activity of TNF-α-converting enzyme (TACE), a membrane-bound disintegrin metalloproteinase).
TNF-α is reported to promote inflammatory cell infiltration by upregulating leukocyte adhesion molecules on endothelial cells, serve as a chemotactic agent for monocytes, and activate phagocyte killing mechanisms. Deficiencies in either TNF-α or its receptors can increase susceptibility to infection by intracellular pathogens. TNF- may also play a role in lymphoid tissue development. Knockout mice lack splenic B cell follicles and the ability to form germinal centers. Other potential physiological roles for TNF-α and its receptors include regulating the differentiation of hematopoietic stem and progenitor cells.
TNF-α has been implicated in a number of pathophysiological processes. It is associated with unregulated pro-inflammatory activity and is thought to be a critical mediator of endotoxin-induced septic shock. Cachexia (or whole body wasting) has also been associated with long-term circulating TNF-α. Other disorders with potential TNF-α involvement include asthma), type 2 diabetes, Crohn’s disease, and rheumatoid arthritis.
GENE ID | 7124 |
SWISS PROT | P01375 |
SYNONYMS | DIF-alpha, TNFA, TNFSF2, TNLG1F, TNF |
Materials Supplied
Kit Components | 96 Wells Quantity/Size |
---|---|
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human TNF-α(8﹡12) | 1 plate |
Human TNF-α Standard lyophilized, 4000 pg/ml upon reconstitution | 2 vials |
Concentrated Biotin-Conjugate anti-human TNF-αmonoclonal 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
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 7pg/mL. |
ASSAY RANGE | 15.6 - 1000 pg/mL |
SPECIFICITY | This assay recognizes both natural and recombinant human 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/Fc, s TNF RII |
Recombinant mouse | TNF-α, TNF-α(truncayed), s TNF RI, s TNF RII IL-4 |
Recombinant rat | TNF-α |
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 BSKH1014.
https://www.myassays.com/bioss-human-tnf-%ce%b1-elisa-kit.assay