Detection of Immobilized Proteins on Nitrocellulose Membranes Using a Biotinylation-Dependent System
作者： Rüdiger Graf , Peter Friedl
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摘要: Proteins are biotinylated after immobilization on nitrocellulose sheets by reaction with a biotinyl-succinimide ester. The biotinyl residues visualized streptavidin-peroxidase-based detection systems either deposition of colored formazan dye or enhanced chemiluminescence (ECL), the latter being 10-fold more sensitive. sensitivity staining procedure is dramatically improved inclusion reporter deposit technique into procedure: initially bound peroxidase generates phenolic radicals from biotinyltyramide, enhancing number in vicinity first biotinylation site. Thus limit lowered to 1 pg protein ECL detection. new method compared silver stain and immunochemical Western blots furthermore its suitability demonstrated for 2-D gel electrophoresis.
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