Aviva Systems Biology TGFA ELISA Kit (Human) (OKBB00252) Product Number OKBB00252 Product Page http://www.avivasysbio.com/tgfa-elisa-kit-human-okbb00252.html Product Name TGFA ELISA Kit (Human) (OKBB00252) Size 96 Tests Gene Symbol TGFA Alias Symbols Transforming growth factor alpha, EGF-like TGF, TGF type 1 NCBI Gene Id 7039 Official Gene Full transforming growth factor alpha Name Target Reference 1. In vivo induction of massive proliferation, directed migration, and differentiation of neural cells in the adult mammalian brain. Proc Natl Acad Sci USA. 2000 Dec 19; 97(26): 14686-91; 2. Brissenden, J. E., Derynck, R., Francke, U. Transforming growth factor alpha gene (TGFA) maps to human chromosome 2 close to the breakpoint of the t(2;8) variant translocation in Burkitt lymphoma. Cytogenet. Cell Genet. 40: 589-only, 1985. 3. Tam, J. P., Scheikh, M. A., Solomon, D. S., Ossowski, L. Efficient synthesis of human type alpha transforming growth factor: its physical and biological characterization. Proc. Nat. Acad. Sci. 83: 8082-8086, 1986. Description of Transforming growth factor alpha (TGF-?) is upregulated in some human cancers. It is Target 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 . TGF? stimulates neural cell proliferation in the adult injured brain.1 Transforming growth factor alpha gene (TGFA) maps to human chromosome 2 close to the breakpoint of the t(2;8) variant translocation in Burkitt lymphoma.2 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 human TGF-alpha showed similar immunoreactivity when compared with rat TGF-alpha. Thus, the 50- amino acid TGF-alpha is likely to be the bioactive principle produced and secreted by tumor cell lines.3 Reconstitution and Store at 4C for frequent use, at -20C for infrequent use. Avoid multiple freeze-thaw Storage cycles (Shipped with wet ice.) Specificity There is cross-reactivates with TGF beta 2, TGF beta 3, TGF beta 5 < 1%. Additional Range: 15.6pg/ml-1000pg/ml Information

5754 Pacific Center Blvd., Suite 201 San Diego, CA 92121 USA | Tel: (858)552-6979 | [email protected] 1 Additional Principle: Aviva’s human TGF? ELISA Kit was based on standard sandwich Information enzyme-linked immune-sorbent assay technology. Human TGF? specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells 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 human TGF? amount of sample captured in plate. Lead Time Domestic: within 1-2 weeks delivery International: 1-2 weeks *** Required Wet/Dry Ice Surcharge will automatically be applied upon checkout for the shipment. See Surcharges Additional Notes: 1. To inspect the validity of experiment operation and the appropriateness of Information sample dilution proportion, pilot experiment using standards and a small number of samples is recommended. 2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case. 3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes. 4. Duplicate well assay is recommended for both standard and sample testing. 5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate. 6. Don’t reuse tips and tubes to avoid cross contamination. 7. To avoid to use the reagents from different batches together. 8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is suggested that the diluted ABC and TMB solution will be pre- warmed in 37C for 30 min before using. Background: Transforming growth factor alpha (TGF-?) is upregulated in some human 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 . TGF? stimulates neural cell proliferation in the adult injured brain. 1 Transforming growth factor alpha gene (TGFA) maps to human chromosome 2 close to the breakpoint of the t(2;8) variant translocation in Burkitt lymphoma.2 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 human TGF-alpha showed similar immunoreactivity when compared with rat TGF-alpha. Thus, the 50- amino acid TGF-alpha is likely to be the bioactive principle produced and secreted by tumor cell lines. Sensitivity < 1 pg/mL Tissue Tool Find tissues and cell lines supported by DNA array analysis to express TGFA. Swissprot Id P01135 Protein Name Protransforming growth factor alpha Protein Accession NP_001093161.1 # Sample Type cell culture supernates, serum, plasma (heparin, EDTA) and human milk RNA Seq Find tissues and cell lines supported by RNA-seq analysis to express TGFA. Nucleotide NM_001099691.2 Accession # 5754 Pacific Center Blvd., Suite 201 San Diego, CA 92121 USA | Tel: (858)552-6979 | [email protected] 2 Kit Component Component Amount Lyophilized recombinant human TGFα standard 10 ng/tube x 2 Anti-human TGFα Antibody Well Plate 96 Wells Sample diluent buffer 30 mL Biotinylated anti- human TGFα antibody 130 uL, dilution 1:100 Antibody diluent buffer 12 mL Avidin-Biotin-Peroxidase Complex (ABC) 130 uL, dilution 1:100 ABC diluent buffer 12 mL TMB color developing agent 10 mL TMB stop solution 10 mL

Kit Reproducibility Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Mean (pg/mL) 118 225 516 126 238 509 St. Dev. 5.43 12.83 16.5 8.95 14.28 27 %CV 4.60 5.70 3.19 7.10 6 5.30 Kit Range 15.6 pg/mL-1,000 pg/mL Kit Duration ~ 3 Hours Kit Principle Aviva Systems Biology TGFA ELISA Kit (Human) (OKBB00252) is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody specific for TGFA has been pre-coated onto 96-wellplate (12 x 8 Well Strips). Standards (E.coli,V40- A89) and test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for TGFA is added, incubated and followed by washing. Avidin- Biotin-Peroxidase Complex is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is visualized through the addition of TMB substrate which is catalyzed by HRP to produce a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm and is quantitatively proportional to the amount of sample Human TGFA captured in well. Detection Method Colorimetric, OD 450 nm CB Replacement sc-1338; sc-27155; sc-36; sc-374433; sc-39423; sc-39424; sc-57447; sc-9043 Species Reactivity Human Application ELISA Predicted Cross-reactivates with TGF?2, TGF?3, TGF?5 <1%. Homology Based on Immunogen Sequence

Image 1: TGFA ELISA standard curve Human TGFA ELISA Kit standard curve

AVIVA SYSTEMS BIOLOGY manufactures and sells quality antibody products covering genome wide proteins. This product is for Research Use Only. Not for diagnostic, human, or veterinary use. Optimal conditions of its use should be determined by end users.

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