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Quantitation of proteins by elution of Coomassie brilliant blue R from stained bands after sodium dodecyl sulfate-polyacrylamide gel electrophoresis
Authors:E H Ball
Institution:1. Medical Oncology Department, Hospital Clínico San Carlos, Madrid, Spain;2. Medical Oncology Department, Hospital Universitario Puerta de Hierro, Madrid, Spain;3. Medical Oncology Department, Centro Integral Oncológico Clara Campal, HM Hospitals, Madrid, Spain;4. Pathology Department, Hospital Clínico San Carlos, Madrid, Spain;1. Key Laboratory of Carbohydrate and Lipid Metabolism Research, College of Life Science and Technology, Dalian University, 10-Xuefu Avenue, Dalian Economical and Technological Development Zone, Liaoning 116622, China;2. School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, Liaoning, China;3. Department of Pharmacology, Institute of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China;1. Laboratory of Microbiology and Virology, Università “Vita-Salute” San Raffaele, Milano, Italy;2. Integral Molecular, Philadelphia, USA;3. Laboratory for Biomolecular Modeling, Institute of Bioengineering, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland;4. Swiss Institute of Bioinformatics, Lausanne, Switzerland;1. College of Horticulture and Landscape Architecture, Northeast Agricultural University, Harbin, 150030, China;2. Key Laboratory of Saline-Alkali Vegetation Ecology Restoration (Northeast Forestry University), Ministry of Education, Harbin, 150040, China;3. College of Life Sciences, Northeast Forestry University, Harbin, 150040, China
Abstract:A simple method for the extraction of Coomassie brilliant blue R from stained protein bands excised from polyacrylamide gels is described. Spectrophotometric measurement of the eluted dye forms the basis of a sensitive assay to quantitate proteins in gels in the range 0.5-10 micrograms. The method requires no unusual equipment and is suitable for measurement of multiple samples. The polypeptide is not extracted and remains available for further analysis. The technique has been applied to three proteins and gels of various acrylamide percentages.
Keywords:
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