Recombinant Human HB-EGF (E. coli-expressed) Protein, CF

R&D Systems | Catalog # 11799-HE

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Key Product Details

Source

E. coli

Accession Number

Q99075.1

Applications

Bioactivity
View all HB-EGF Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human HB-EGF protein
Asp63-Leu148

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Asp63

Predicted Molecular Mass

9.7 kDa

SDS-PAGE

8-12 kDa, under reducing conditions.

Activity

Measured in a cell proliferation assay using Balb/3T3 mouse embryonic fibroblast cells. Rubin, J.S. et al. (1991) Proc. Natl. Acad. Sci. USA 88:415.
The ED50 for this effect is 0.500-5.00 ng/mL.

Scientific Data Images for Recombinant Human HB-EGF (E. coli-expressed) Protein, CF

Recombinant Human HB‑EGF Protein Bioactivity.

Recombinant Human HB‑EGF Protein (Catalog # 11799-HE) induces cell proliferation in Balb/3T3 mouse embryonic fibroblast cells.

Recombinant Human HB‑EGF Protein SDS-PAGE.

2 μg/lane of Recombinant Human HB‑EGF Protein (Catalog # 11799-HE) was resolved with SDS-PAGE under reducing (R) condition and visualized by Coomassie® Blue staining, showing bands at 8-12 kDa.

Formulation, Preparation, and Storage

11799-HE
Formulation Lyophilized from a 0.2 μm filtered solution in NaPO4 and NaCl.
Reconstitution Reconstitute at 100 μg/mL in water.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: HB-EGF

Human HB-EGF (Heparin-Binding EGF-like growth factor) is a 12-16 kDa member of the EGF family of peptide growth factors (1-3). Also known as the DTR (diphtheria toxin receptor), it is further classified as a group 2 ErbB ligand based on its ability to activate both the EGF/ErbB1 and ErbB4 receptors (4, 5). HB-EGF is synthesized as a 208 amino acid (aa) type I transmembrane preproprecursor (1, 6). It contains a 19 aa signal sequence, a 43 aa prosegment, an 86 aa mature region (aa 63-148), an 11 aa juxtamembrane cleavage peptide, a 24 aa transmembrane segment, and a 25 aa cytoplasmic tail (aa 184-208). As an integral membrane protein, HB-EGF is expressed as a 19-27 kDa protein in mammalian cells (7-9). The variability in molecular weight (MW) is attributed to heterogeneity in glycosylation and/or the utilization of multiple proteolytic cleavage sites during maturation. Mature HB-EGF is a soluble peptide that arises from proteolytic processing of the transmembrane form. It possesses an EGF-like domain between aa 104-144, and a heparin-binding motif between aa 93‑113. Although the aa range for "mature" HB-EGF is typically stated to be Asp63-Leu148, potential N-terminal start (cleavage) sites also exist at Gly32, Arg73, Val74, Ser77 and Ala82 (8, 10-12). Thus, differential processing (in part) likely accounts for the 16-23 kDa range in MW noted for mammalian-derived mature HB-EGF. Proteases suggested to contribute to HB-EGF processing include TACE, MMP-3 and -7, ADAM-17 and ADAM-12 (11, 13-16). When expressed recombinantly in E.coli, HB-EGF (aa 73-148) runs at 14 kDa in SDS-PAGE; when expressed in Baculovirus, HB-EGF (aa 63-148, 77-148 and 32-148) runs at 18 kDa, 15 kDa, and 19 kDa respectively (8, 12, 17). Over aa 63-148, human HB-EGF- shares 76% and 73% aa sequence identity with rat and mouse HB-EGF, respectively (1, 18). Cells known to express HB-EGF include bronchial epithelium (19), visceral and vascular smooth muscle (20, 21), CD4+ T cells (22), cardiac muscle (23), glomerular podocytes (24), keratinocytes (13) and IL-10-secreting regulatory macrophages (25). As noted earlier, HB-EGF is known to bind to both 170 kDa EGFR and 180 kDa ErbB4, and through heterodimerization, ErbB2 (13, 26). Activity associated with ErbB4 binding appears to be limited to non-mitogenic actions, while EGFR binding induces both mitogenic and non-mitogenic activity.

References

  1. Higashiyama, S et al. (1991) Science 251:936.
  2. Schneider, MR & E. Wolf (2009) J. Cell. Physiol. 218:460.
  3. G.V. Sherbet (2011) The Epidermal Growth Factor (EGF) Family in Growth Factors and Their Receptors in Cell Differentiation, Cancer and Cancer Therapy, Pages 173-198. Elsevier, NY.
  4. Iwamoto, R & E. Mekada (2000) Cytokine Growth Factor Rev. 11:335.
  5. Miyata, K. et al. (2012) Anticancer Res. 32:2347.
  6. SwissProt:Q99075.
  7. Raab, G. et al. (1994) Biochem. Biophys. Res. Commun. 204:592.
  8. Nakagawa, T. et al. (1996) J. Biol. Chem. 271:30858.
  9. Higashiyama, S. et al. (1995) J. Cell Biol. 128:929.
  10. Higashiyama, S. et al. (1992) J. Biol. Chem. 267:6205.
  11. Hinkle, C.L. et al. (2004) J. Biol. Chem. 279:24179.
  12. Ono, M. et al. (1994) J. Biol. Chem. 269:31315.
  13. Nanba, D. & S. Higashiyama (2004) Cytokine Growth Factor Rev. 15:13.
  14. Cheng, K. et al. (2007) Biochem. Pharmacol. 73:1001.
  15. Suzuki, M. et al. (1997) J. Biol. Chem. 272:31730.
  16. Asakura, M. et al. (2002) Nat. Med. 8:35.
  17. Marikovsky, M. et al. (1993) Proc. Natl. Acad. Sci. USA 90:3889.
  18. Abraham, J.A. et al. (1993) Biochem. Biophys. Res. Commun. 190:125.
  19. Tschumperlin, D.J. et al. (2004) Nature 429:83.
  20. Park, J.M. et al. (1998) Am. J. Physiol. 275:C1247.
  21. Miyagawa, J. et al. (1995) J. Clin. Invest. 95:404.
  22. Blotnick, S. et al. (1994) Proc. Natl. Acad. Sci. USA 91:2890.
  23. Iwamoto, R. et al. (2003) Proc. Natl. Acad. Sci. USA 100:3221.
  24. Bollee, G. et al. (2011) Nat. Med. 17:1242.
  25. Edwards, J.P. et al. (2009) J. Immunol. 182:1929.
  26. Elenius, K. et al. (1997) EMBO J. 16:1268.

Long Name

Heparin Binding EGF-like Growth Factor

Alternate Names

Dtr, Dts, HBEGF, Hegfl

Entrez Gene IDs

1839 (Human); 15200 (Mouse)

Gene Symbol

HBEGF

UniProt

Q99075.1

Additional HB-EGF Products

Product Documents for Recombinant Human HB-EGF (E. coli-expressed) Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human HB-EGF (E. coli-expressed) Protein, CF

For research use only

Related Research Areas

EGF Family

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