Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Transforming Growth Factor α (hTGF-α) #5495

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Source

Recombinant human TGF-α (hTGF-α) Val40-Ala89 (Accession #NP_003227) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Based on amino acid sequencing, greater than 50% of recombinant hTGF-α has a Met on the amino-terminal Val40 (MVVSH) and has a calculated MW of 5,683. The remainder starts at Val40 (VVSHF). 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 hTGF-α. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hTGF-α was determined in a MCF-10A cell proliferation assay. The ED50 of each lot is between 0.25-1.0 ng/ml.

Coomassie Gel

Coomassie Gel

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

Bioactivity

Bioactivity

The proliferation of MCF-10A cells treated with increasing concentrations of hTGF-α was assessed. After 24 hour treatment, cells were labeled with BrdU for 4 hours. BrdU incorporation was determined by ELISA and the OD450-OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells untreated or treated with hTGF-α for 10 minutes, using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb #4407 (upper) and EGF Receptor Antibody #2232 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hTGF-α.

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

Background

TGF-α is a member of the EGF family of proteins and is expressed by epithelial cells, keratinocytes, macrophages, and many tumor-derived cell lines (1-4). Soluble TGF-α is produced by proteolytic cleavage of a transmembrane precursor (5). TGF-α promotes cell proliferation and differentiation of a diverse array of cell types including cancer and neural cells (1,4,6). TGF-α binds to EGFR/ErbB1 and induces receptor dimerization and tyrosine kinase activation (7). TGF-α activates the MAPK pathways and involves Stat3 and Stat5 (1,6). Increased levels of TGF-α expression are associated with many cancers including breast cancer (1,8,9).

  1. Humphreys, R.C. and Hennighausen, L. (2000) Oncogene 19, 1085-91.
  2. Rappolee, D.A. et al. (1988) Science 241, 708-12.
  3. Xu, X. et al. (1999) Oncogene 18, 5554-62.
  4. Ciana, P. et al. (2003) J Biol Chem 278, 31737-44.
  5. Higashiyama, S. et al. (2008) Cancer Sci 99, 214-20.
  6. Song, J.I. and Grandis, J.R. (2000) Oncogene 19, 2489-95.
  7. Citri, A. and Yarden, Y. (2006) Nat Rev Mol Cell Biol 7, 505-16.
  8. Révillion, F. et al. (2008) Ann Oncol 19, 73-80.
  9. Masago, K. et al. (2008) Cancer Sci 99, 2295-301.

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

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