Description
Tumor necrosis factor α (TNFα or TNF) is secreted by macrophages in response to inflammation, infection, and cancer. Rat Tumor Necrosis Factor (TNF) and Lymphotoxin (TNFβ) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology. TNFα is produced by monocytes, which can stimulate endothelial cells to produce the multi-lineage growth factor granulocyte-macrophage colony-stimulating factor (GM-CSF) and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells, but has no effect on normal cells. Rat TNF has been purified to apparent homogeneity as a 17.3kDa protein from HL-60 leukemia cells and has shown cytotoxic and cytostatic activities against various rat tumor cell lines. The Rat TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNFα has been mapped to human chromosome 6. The standard product used in this kit is recombinant rat TNFα, consisting of 157 amino acids with a molecular mass of 17kDa. ScienCell's rat TNFα ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. Rat TNFα-specific monoclonal antibodies are pre-coated onto 8 x 12 strips. The rat-specific detection polyclonal antibodies are biotinylated. The test samples and biotinylated detection antibodies are subsequently added to the wells and then washed with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with PBS or TBS buffer. HRP substrate TMB is used to visualize HRP enzymatic reaction. TMB is catalyzed by HRP to produce a blue color product that changes to yellow after adding acidic stop solution. The intensity of yellow is proportional to the amount of rat TNFα in the sample that is captured on the strips