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Description
Human HB-EGF was initially identified as a protein of 22 kD secreted by macrophage-like U937 cells. It belongs to the EGF family of proteins that includes EGF, TGF-α, heparin-binding EGF like-growth factor (HB-EGF), epigen, epiregulin, betacellulin, neuroregulin, and tomoregulin. All the EGF family members are synthesized as type I membrane protein precursors, which can undergo proteolytic cleavage at the plasma membrane to release a mature soluble ectodomain.  It has been suggested that various metalloproteinases participate in the shedding of HB-EGF, such as MMP-3, MMP-7, ADAM9, ADAM10, ADAM12, and ADAM17. The ectodomain shedding is stimulated by phorbol esters, calcium ionophore, lysophosphatidic acid, and IL-1β. In addition, nardilysin, a metalloendopeptidase of the M16 family, binds HB-EGF and enhances its shedding through activation of TACE (ADAM17). The membrane-anchored HB-EGF acts in cell-cell and cell-extracellular matrix interactions; in addition, the membrane HB-EGF is the receptor for diphtheria toxin that can be internalized and induce apoptotic death. HB-EGF plays a crucial role in cardiac valvulogenesis; newborn knockout mice have malformed semilunar and atrioventricular heart valves and poorly differentiated lungs. HB-EGF is overexpressed in several types of cancer such as pancreatic carcinoma, ovarian cancer, and gastric carcinoma. Also, HB-EGF is involved in cancer metastasis and invasion of ovarian cancer, head and neck cancer, and thyroid carcinoma cells.
Specifications

General

SourceHuman HB-EGF-like growth factor, amino acids (Asp63-Leu148) (Accession# NM_001945), was expressed in E. coli.
Molecular MassThe 86 amino acid recombinant protein has a predicted molecular mass of approximately 9.7 kD. The DTT-reduced and non-reduced protein migrate at approximately 13 kD and 16 kD respectively by SDS-PAGE. The predicted N-terminal amino acid is Asp.
Purity>95%, as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in PBS, pH 7.2.
Endotoxin LevelLess than 0.01 ng per µg cytokine as determined by the LAL method.
Concentration10 and 25 µg sizes are bottled at 100 µg/mL.
Storage & HandlingUnopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
ActivityRecombinant human HB‐EGF induces proliferation of fetal mouse fibroblast cells, BALB/3T3 in a dose dependent manner. ED50 for this effect of 0.5 - 5.0 ng/mL.
ApplicationBioassay
Application NotesBioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
DistributionMonocytes, macrophages, T cells, neutrophils, eosinophils, myeloid leukemia blasts, myeloma cells, vascular smooth muscle cells, endothelial cells, and normal or neoplastic epithelial cells.
FunctionHB-EGF is associated with proliferation and differentiation of stromal cells, apoptosis, and morphogenesis. It participates in inflamation, wound healing, atherosclerotic plaque progression, and cardiac valvulogenesis. TNF-α and IL-1β induces HB-EGF in endothelial cells.
InteractionFibroblast, smooth muscle cells, monocytes/macrophages, endothelial cells, astrocytes, keratinocytes, normal and neoplastic epithelial cells.
Ligand/ReceptorEGFR (ErbB1) and ErbB4; also binds to heparan-sulphate proteoglycans.
Cell TypeEmbryonic Stem Cells, Hematopoietic stem and progenitors, Mesenchymal Stem Cells, Neural Stem Cells
Biology AreaCell Biology, Neuroscience, Stem Cells, Synaptic Biology
Molecular FamilyCytokines/Chemokines, Growth Factors
Antigen References1. Higashiyama S, et al. 1991. Science 251:936. 2. Hirata M, et al. 2001. Biochem. Biophys. Res. Commun. 283:915. 3. Jackson LF, et al. 2003. EMBO J. 22:2704. 4. Nishi E, et al. 2006. J. Biol. Chem. 281:31164. 5. Yotsumoto F, et al. 2008. Biochem. Biophys. Res. Commun. 365:555. 6. Ota I, et al. 2013. Oncol Rep. 30:1593.
Gene ID1839
UniProtView information about HB-EGF on UniProt.org
Regulatory StatusRUO
Other NamesHeparin-binding EGF-like growth factor, HB-EGF-like growth factor
Biolegend 552002 Recombinant Human HB-EGF (carrier-free) | IRIGHT