Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Epigen #5493

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Source

Recombinant human Epigen (hEpigen) Ala24-Ala104 (Accession #Q6UW88-1) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Recombinant hEpigen does not have a Met on the amino terminus and has a calculated MW of 9,087. DTT-reduced and non-reduced protein migrate as 8 kDa polypeptides with non-reduced protein having slightly greater mobility due to intramolecular cystines. The expected amino-terminal AVTVT of recombinant hEpigen was verified by amino acid sequencing.

Purity

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

Bioactivity

The bioactivity of recombinant hEpigen was determined in a MCF-10A cell proliferation assay. The ED50 of each lot is between 2-30 ng/ml.

Coomassie Gel

Coomassie Gel

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

Bioactivity

Bioactivity

The proliferation of MCF-10A cells treated with increasing concentrations of hEpigen 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 hEpigen for 10 minutes, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hEpigen.

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 hEpigen. Carrier free: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl.

Background

Epigen was identified as an EGF family member by amino acid sequence homology (1). The sequence was obtained from mouse keratinocyte mRNA. Epigen is expressed in normal human mammary tissue and in some prostate and breast carcinomas (2). Epigen binding to known EGF receptors is with lower affinity when compared to EGF. However, Epigen appears to be superior in inducing cell proliferation (2). Epigen induces responses in MDA-MB-468 breast carcinoma cells. Epigen also has effects on prostate cells and vascular endothelial cells (2). Epigen binds to ErbB1/Her1 (2). Binding causes receptor homodimerization and heterodimerization with other Erb/Her receptors (2).

  1. Strachan, L. et al. (2001) J Biol Chem 276, 18265-71.
  2. Kochupurakkal, B.S. et al. (2005) J Biol Chem 280, 8503-12.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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