PF078

Heparin Binding Epidermal Growth Factor, Human, Recombinant, S. frugiperda

• Synonym(s): HB-EGF
• UNSPSC Code: 12352202

Properties

• Assay: ≥97% (SDS-PAGE)
• Quality Level: 100
• form: solid
• manufacturer/tradename: Calbiochem^®
• storage condition: OK to freeze; avoid repeated freeze/thaw cycles
• impurities: ≤1 EU/μg Endotoxins (EU/μg cytokine)
• shipped in: wet ice
• storage temp.: −20°C

Description

General description

Recombinant, human heparin binding epidermal growth factor expressed in S. frugiperda insect cells. The mature HB-EGF is a human recombinant protein whose DNA sequence encoding the N-terminal 148 amino acid residues of human HB-EGF precursor was expressed in Sf 21 insect cells using a baculovirus expression system. The mature recombinant protein (86 amino acids) generated by the removal of the 62 amino acid residue signal and propeptide has a predicted molecular mass of ~9.5 kDa. The recombinant protein is heterogeneously O-glycosylated and migrates as an ~12 kDa protein in SDS-PAGE. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a ~22 kDa O-glycosylated protein that is a potent mitogen and chemoattractant for vascular smooth muscle cells, fibroblasts, and epithelial cells but not endothelial cells. The natural protein has an apparent molecular mass of ~19-23 kDa and exists in multiple forms as a result of heterogeneous O-glycosylation and/or N-terminal truncation. HB-EGF is synthesized as a membrane-anchored precursor (proHB-EGF) that is proteolytically cleaved to release the soluble mature growth factor. The two forms are active as juxtacrine and paracrine/autocrine growth factors respectively. Recent studies show MMP-3 as one of the enzymes that cleaves HB-EGF implicating a role for MMP-3 in the regulation of HB-EGF from being a juxtacrine to a paracrine/autocrine factor. HB-EGF activates two EGF receptor subtypes, HER1/ErbB1, and HER4 and binds to heparan sulfate proteoglycan. The proHB-EGF is postulated as the high affinity receptor for diphtheria toxin (DT). Recent evidence suggests that the coexpression of proHB-EGF and CD9 on macrophages may strongly promote the development of atherosclerosis by a juxtacrine mechanism. Transcription of HB-EGF can be induced in vascular endothelial cells as well as aortic smooth muscle cells which may also play an important role in the pathogenesis of atherosclerosis. Useful for proliferation studies.

Recombinant, human heparin binding epidermal growth factor expressed in S. frugiperda insect cells. The mature HB-EGF is generated by the removal of the 62 amino acid (~9.5 kDa) signal. Protein is heterogeneously O-glycosylated and migrates as an ~12 kDa protein in SDS-PAGE. Titrate the protein for optimal results in individual systems.

Biochem/physiol Actions

EC₅₀ of 2-5 ng/ml as measured by its ability to stimulate ³H-thymidine incorporation in the EGF-responsive fibroblast cell line, Balb/3T3

Warning

Toxicity: Standard Handling (A)

Physical form

Lyophilized from a sterile-filtered PBS solution, 50 µg of BSA/µg of cytokine, pH 7.4.

Reconstitution

Reconstitute in PBS to ≥1 &microg/ml containing ≥0.1%HSA or BSA.

Other Notes

Davis-Fleischer, K.M. and Besner, G.E. 1998. Front. Biosci.3, D288.
Ouchi, N., et al. 1997. Biochem. J.328, 923.
Raab, G. and Klagsbrun, M. 1997. Biochim. Biophys. Acta1333, F179.
Suzuki, M., et al. 1997. J. Biol. Chem.272, 31730.
Nakata, A., et al. 1996. Circulation94, 2778.
Miyagawa, J., et al. 1995. J. Clin. Invest.95, 404.
Higashiyama, S., et al. 1991. Science251, 936.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 3