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Open Access Highly Accessed Methodology article

Highly sensitive and specific bioassay for measuring bioactive TGF-β

Ina Tesseur1, Kun Zou13, Elisabeth Berber1, Hui Zhang1 and Tony Wyss-Coray12*

Author Affiliations

1 Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA

2 GRECC, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA

3 Department of Alzheimer's Disease Research, National Center for Geriatrics and Gerontology, Obu, Aichi 474-8522, Japan

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BMC Cell Biology 2006, 7:15  doi:10.1186/1471-2121-7-15

Published: 20 March 2006

Abstract

Background

Transforming Growth Factor-β (TGF-β) regulates key biological processes during development and in adult tissues and has been implicated in many diseases. To study the biological functions of TGF-β, sensitive, specific, and convenient bioassays are necessary. Here we describe a new cell-based bioassay that fulfills these requirements.

Results

Embryonic fibroblasts from Tgfb1-/- mice were stably transfected with a reporter plasmid consisting of TGF-β responsive Smad-binding elements coupled to a secreted alkaline phosphatase reporter gene (SBE-SEAP). Clone MFB-F11 showed more than 1000-fold induction after stimulation with 1 ng/ml TGF-β1, and detected as little as 1 pg/ml TGF-β1. MFB-F11 cells were highly induced by TGF-β1, TGF-β2 and TGF-β3, but did not show induction with related family members activin, nodal, BMP-2 and BMP-6 or with trophic factors bFGF and BDNF. MFB-F11 cells can detect and quantify TGF-β in biological samples without prior enrichment of TGF-βs, and can detect biologically activated TGF-β in a cell co-culture system.

Conclusion

MFB-F11 cells can be used to rapidly and specifically measure TGF-β with high sensitivity.