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J Biochem Biophys Methods. 2005 Jul 29;64(1):76-81.

Detection of neuronal growth inhibitory factor (metallothionein-3) in polyacrylamide gels and by Western blot analysis.

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  • 1Institute of Biochemistry, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.

Abstract

Neuronal growth inhibitory factor (GIF) is a small cysteine-rich metal binding protein downregulated in Alzheimer's disease. The protein belongs to the superfamily of metallothioneins (MTs) and was classified as MT-3. Although first identified as a brain specific protein, several reports now indicate a substantially broader expression pattern. However, currently available detection methods for MT-3 show low sensitivity in gel electrophoresis and Western blot. We have developed a fast and sensitive method for MT-3 detection in SDS-PAGE (detection limit approximately 10 ng) and Western blot (detection limit approximately 1 ng). The method is based on the chemical modification of cysteine residues with the dye monobromobimane and an improved blotting protocol.

PMID:
15982745
DOI:
10.1016/j.jbbm.2005.05.005
[PubMed - indexed for MEDLINE]
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