Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Epiregulin #5494

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Source

Recombinant human Epiregulin (hEpiregulin) Val63-Leu108 (Accession #NP_001423) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Based on amino acid sequencing, greater than 50% of recombinant hEpiregulin has a Met on the amino-terminal Val63 (MVSIT) and has a calculated MW of 5,406. The remainder starts at Val63 (VSITK). DTT-reduced and non-reduced protein migrate as 5 kDa polypeptides.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hEpiregulin. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hEpiregulin was determined in a MCF 10A cell proliferation assay. The ED50 of each lot is between 0.10-0.85 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hEpiregulin was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hEpiregulin and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of MCF 10A cells treated with increasing concentrations of hEpiregulin was assessed. After 24 hr treatment, cells were labeled with BrdU for 4 hr. BrdU incorporation was determined by ELISA and the OD450-OD690 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF 10A cells, untreated or treated with hEpiregulin for 10 minutes, using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb #4407 (upper) and EGF Receptor (C74B9) Rabbit mAb #2646 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hEpiregulin.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl and 20 μg BSA per 1 μg hEpiregulin. Carrier free: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl.

Background

Epiregulin is a member of the EGF family that is highly expressed in many cancer-specific cells, cells of the placenta and macrophages (1,2). Epiregulin induces keratinocyte and corneal epithelial cell proliferation, oocyte maturation and may function in blastocyst implantation during pregnancy (3-5). Active soluble epiregulin is produced by proteolytic processing of a transmembrane precursor. Epiregulin is the only EGF family member known to bind to both the ErbB1/HER1 and ErbB4/HER4 EGF receptors (6). Epiregulin binding induces receptor homodimerization or heterodimerization with other Erb/Her receptors. Epiregulin activates the ERK1/2, and PI3K/Akt pathways (1,3).

  1. Toyoda, H. et al. (1997) Biochem J 326 ( Pt 1), 69-75.
  2. Révillion, F. et al. (2008) Ann Oncol 19, 73-80.
  3. Morita, S. et al. (2007) Mol Vis 13, 2119-28.
  4. Shirakata, Y. et al. (2000) J Biol Chem 275, 5748-53.
  5. Song, H. et al. (2000) Mol Endocrinol 14, 1147-61.
  6. Riese, D.J. et al. (1998) J Biol Chem 273, 11288-94.

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