Recombinant Human HB-EGF

Heparin-binding EGF-like growth factor (HB-EGF), also known as DTR and DTS, is a heparin binding, O-glycosylated protein and a member of the EGF family of proteins. It is a potent mitogen and chemoattractant for vascular smooth muscle cells, fibroblasts and epithelial cells but not endothelial cells. HB-EGF is synthesized as a membrane-anchored precursor that is proteolytically cleaved to release the soluble mature growth factor. The two forms are active as juxtacrine and paracrine/autocrine growth factors respectively. HB-EGF activates two EGF receptor subtypes, HER1/ErbB1 and HER4 and binds to heparin sulfate proteoglycan. HB-EGF is expressed in numerous cell types and tissues, including vascular endothelial cells and SMC, macrophages, skeletal muscle, keratinocytes and certain tumor cells. It has been shown to play a role in wound healing, cardiac hypertrophy and heart development and function (2). HB-EGF contributes to cell adhesion, invasion, and angiogenesis, which are integral to transcoelomic metastasis in ovarian cancer (3). Production of HB-EGF in vascular smooth muscle cells is induced by angiotensin, thus it may have an important autocrine role in the proliferation of these cells in vascular diseases such as atherosclerosis and hypertension. HB-EGF has been shown to interact with NRD1, zinc finger and BTB domain-containing protein 16 and BAG1.

The biological activity of Human HB-EGF was determined by its ability to stimulate proliferation in a mouse fibroblast cell line, Balb/3T3.⁴ The expected ED₅₀ for this effect is typically 0.15 - 0.75 ng/ml. The cell number is assessed in a fluorometric assay using the redox sensitive dye, Resazurin.

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Asp63 (major), Gly32, Glu24

Lyophilized

The predicted molecular weight of Recombinant Human HB-EGF is Mr 9.5 kDa. However, this recombinant protein is heterogenously O-glycosylated and the actual molecular weight as observed by migration on SDS-PAGE is Mr 11-20 kDa.

9.5

This recombinant protein was 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 with no calcium, magnesium, or preservatives present.

This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.

Next Day Ambient

1. Higashiyama, S. et al. (1993) J. Cell Biol. 122:933
2. Higashiyama, S. et al. (2004) Cytokine Growth Factor Rev. 15:13
3. Miyamoto, S. et al. (2008) Mol. Cancer Ther. 7:3441
4. Rubin, JS. et al. (1991) Proc. Natl. Acad. Sci. USA 88:415