Transforming growth factor alpha(TGF-alpha) is upregulated in some Equine cancers. It is produced in macrophages, brain cells, and keratinocytes, and induces epithelial development. It is closely related to EGF, and can also bind to the EGF receptor with similar effects. TGFalpha stimulates neural cell proliferation in the adult injured brain. Transforming growth factor alpha gene(TGFA) maps to Equine chromosome 2 close to the breakpoint of the t(2,8) variant translocation in Burkitt lymphoma. Synthetic TGF-alpha was as active as murine epidermal growth factor in binding to the epidermal growth factor receptor and in stimulation of anchorage-dependent and of anchorage-independent growth of normal indicator cells in culture. Synthetic TGF-alpha stimulated plasminogen activator production in A 431 and HeLa cells, the stimulation was similar to that induced by epidermal growth factor. Furthermore, synthetic Equine TGF-alpha showed similar immunoreactivity when compared with Equine TGF-alpha. Thus, the 50aaTGF- alpha is likely to be the bioactive principle produced and secreted by tumor cell lines. Sample Type: Cell culture supernates, serum, human milk, plasma (heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Equine TGF alpha. Sensitivity: <1pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Equine TGF alpha. There is cross-reactivity with TGF beta 2, TGF beta 3, TGF beta 5 (<1%). Test Principle: This Equine TGF Alpha BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Equine TGF Alpha has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli,V40-A89) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Equine TGF Alpha is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Equine TGF Alpha amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Equine TGF alpha antibody, 1x plate Lyophilized recombinant Equine TGF alpha standard, 2x10ng/tube Biotinylated anti-Equine TGF alpha antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
