Mouse TNF-alpha Antibody Summary
Leu80-Leu235, with an N-terminal Met & Ser84-Leu235
Accession # P06804
Applications
This antibody functions as an ELISA capture antibody when paired with Goat Anti-Mouse TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-410-NA).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Mouse TNF-alpha DuoSet ELISA Kit (Catalog # DY410) for convenient development of a sandwich ELISA or the Mouse TNF-alpha Quantikine ELISA Kit (Catalog # MTA00B) for a complete optimized ELISA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
Mouse TNF‑ alpha ELISA Standard Curve. Recombinant Mouse TNF-a protein was serially diluted 2-fold and captured by Rat Anti-Mouse TNF-a Monoclonal Antibody (Catalog # MAB4102) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-410-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: TNF-alpha
Tumor necrosis factor alpha (TNF-alpha, TNF- alpha, TNFA ), 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, immune system development, apoptosis, and lipid metabolism. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Mouse TNF-alpha consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD). Within the ECD, mouse TNF-alpha shares 94% aa sequence identity with rat and 70%-77% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.
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