Recombinant Human HGF (NK1), Animal-Free Protein

HGF NK1 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. Cells were treated in duplicate with a serial dilution of HGF for 6 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. EC50 = 1.5 ng/ml (72.9 pM).*Promega pGL4.33[luc2P/SRE/Hygro] #E1340

HGF NK1 migrates as major band at 20 kDa in non-reducing conditions and 18 kDa upon reduction.Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+DTT, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250. NB reduced samples were not boiled as the protein is sensitive to high temperatures, which causes degradation.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human HGF (NK1), Animal-Free Protein Summary

• Details of Functionality: No significant difference between EC₅₀ of reference and test lots
• Source: E. coli-derived human HGF protein
• Accession #: P14210.2
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 20 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: Monomeric HGF (NK1) protein only

Packaging, Storage & Formulations

• Storage: Store lyophilized protein between -20 and -80 °C until the date of expiry. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from acetonitrile/TFA
• Reconstitution Instructions: Resuspend in 10mM HCl at >100 µg/ml, prepare single use aliquots, add carrier protein if desired.

Notes

The above product was manufactured, tested and released by R&D System's contract manufacturer, Qkine Ltd, at 1 Murdoch House, Cambridge, UK, CB5 8HW. The product is for research use only and not for the diagnostic or theraputic use.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human HGF (NK1), Animal-Free Protein

• deafness, autosomal recessive 39
• DFNB39
• EC 3.4.21
• EC 3.4.21.7
• fibroblast-derived tumor cytotoxic factor
• F-TCF
• hepatocyte growth factor (hepapoietin A; scatter factor)
• Hepatopoeitin-A
• Hepatopoietin A
• HGF
• HGFB
• HPTA
• HPTAhepatocyte growth factor
• lung fibroblast-derived mitogen
• Scatter factor
• SF
• SFhepatopoeitin-A

Background

HGF, also known as scatter factor and hepatopoietin A, is a pleiotropic protein in the plasminogen subfamily of S1 peptidases. It is a multidomain molecule that includes an N-terminal PAN/APPLE-like domain, four Kringle domains, and a serine proteinase-like domain that has no detectable protease activity (1-5) Human HGF is secreted as an inactive 728 amino acid (aa) single chain propeptide. It is cleaved after the fourth Kringle domain by a serine protease to form bioactive disulfide-linked HGF with a 60 kDa alpha and 30 kDa beta chain. Alternate splicing generates human HGF isoforms that lack the proteinase-like domain and different numbers of the Kringle domains. Human HGF shares 91%-94% aa sequence identity with bovine, canine, feline, mouse, and rat HGF. HGF binds heparan-sulfate proteoglycans and the widely expressed receptor tyrosine kinase, HGF R/c-MET (6, 7). HGF-dependent c-MET activation is implicated in the development of many human cancers (8). HGF regulates epithelial morphogenesis by inducing cell scattering and branching tubulogenesis (9, 10). HGF induces the up-regulation of integrin alpha 2 beta 1 in epithelial cells by a selective increase in alpha 2 gene transcription (11). This integrin serves as a collagen I receptor, and its blockade disrupts epithelial cell branching tubulogenesis (11, 12). HGF can also alter epithelium morphology by the induction of nectin-1 alpha  ectodomain shedding, an adhesion protein component of adherens junctions (13). In the thyroid, HGF induces the proliferation, motility, and loss of differentiation markers of thyrocytes and inhibits TSH-stimulated iodine uptake (14). HGF promotes the motility of cardiac stem cells in damaged myocardium (15).

1. Karihaloo, A. et al. (2005) Nephron Exp. Nephrol. 100:e40.
2. Hammond, D.E. et al. (2004) Curr. Top. Microbiol. Immunol. 286:21.
3. Rosario, M. and Birchmeier, W. (2004) Dev. Cell 7:3.
4. Lesk, A.M. and Fordham, W.D. (1996) J. Mol. Biol. 258:501.
5. Nakamura, T. et al. (1989) Nature 342:440.
6. Mizuno, K., et al. (1994) J. Biol. Chem. 269:1131.
7. Gheradi, E. et al. (2003) Proc. Natl. Acad. Sci. 100:12039.
8. Corso, S. et al. (2005) Trends Mol. Med. 11:284.
9. Maeshima, A. et al. (2000) Kid. Int. 58:1511.
10. Montesano, R. et al. (1991) Cell 67:901.
11. Chiu, S-J. et al. (2002) J. Biomed. Sci. 9:261.
12. Saelman, E.U.M. et al. (1995) J. Cell Sci. 108:3531.
13. 13. Tanaka, Y. et al. (2002) Biochem. Biophys. Res. Commun. 299:472.
14. 14. Mineo, R. et al. (1994) Endocrinology 145:4355.
15. 15. Urbanek, K. et al. (2005) Circ. Res. 97:663.

Bioinformatics

• Uniprot:
  • P14210.2()