Transforming growth factor alpha(TGF-alpha) is upregulated in some Porcine 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 Porcine 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 Porcine TGF-alpha showed similar immunoreactivity when compared with Porcine TGF-alpha. Thus, the 50aa TGF- 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 Porcine TGF alpha. Sensitivity: <1pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Porcine TGF alpha. There is cross-reactivity with TGF beta 2, TGF beta 3, TGF beta 5 (<1%). Test Principle: The Porcine TGF Alpha BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Porcine 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 Porcine 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 Porcine TGF Alpha amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Porcine TGF alpha antibody, 1x plate Lyophilized recombinant Porcine TGF alpha standard, 2x10ng/tube Biotinylated anti-Porcine 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.
