Detection of neuronal growth inhibitory factor (metallothionein-3) in polyacrylamide gels and by Western blot analysis.
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.
Meloni
Gabriele
G
Institute of Biochemistry, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
Knipp
Markus
M
Vasák
Milan
M
D000544:Alzheimer Disease / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D001643:Bridged Bicyclo Compounds / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D000090023:Metallothionein 3; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity
D001643:Bridged Bicyclo Compounds; D000090023:Metallothionein 3; D009419:Nerve Tissue Proteins; C028381:monobromobimane