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Updated: Jun 21, 2026
Western Blotting: Sample Preparation to Detection
Published on: October 14, 2010
Facile detection of proteins on a solid-phase membrane by direct binding of dextran-based luminol-biotin
Huan Zhang1, Takayuki Shibata, Tomasz Krawczyk
1Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14, Bunkyo-Machi, Nagasaki 852-8521, Japan.
Abstract:
Facile and non-radioactive methods are desired for the sensitive detection and quantification of various proteins. Herein we describe a novel chemiluminescence (CL)-detection method of particular proteins based on direct binding of a dextran-luminol-biotin (DLB) CL polymer to the proteins on a poly(vinylidene difluoride) membrane. Among 32 kinds of the proteins screened, several proteins such as drug-metabolizing enzymes, cytochrome p450 (CYP)1A2, CYP2E1, and CYP3A4 had the ability to bind directly to the DLB polymer. The binding site in the polymer was owing to the framework of the modified dextran, which underwent oxidation and reduction procedures. This interaction might be the comprehensive effect of both electrostatic interaction and steric complementarities. CL intensity of the proteins detected by the polymer could be further enlarged by the mediation of avidin. The proposed CL-imaging method possesses potential as a rapid, facile, inexpensive and selective detection of the proteins.
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