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2005•Journal of Shantou University Medical CollegeRequires access

A Modified Electrophoretic Mobility Shift Assay For Nuclear Factor-kappa B

Ling‐Fei Wu

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Abstract

Objective:To establish a fast and simple method of electrophoretic mobility shift assay(EMSA)for the identification of DNA-binding protein nuclear factor-kappa B(NF-κB).Methods:NF-κB protein was extracted and its concentration was determined.DNA probe was labeled by ()~(32)P.The probe was conjugated with NF-κB protein together and underwent electrophoresis.The X-ray film was exposed and autoradiography was obtained.Results:By using this method,highly distinct bands and clear background in the picture were obtained and the activity of NF-κB could be detected by eyes.Conclusion:The modified EMSA is more simple,feasible,and highly sensitive method for detection of DNA-binding proteins.

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What this paper is about

Objective:To establish a fast and simple method of electrophoretic mobility shift assay(EMSA)for the identification of DNA-binding protein nuclear factor-kappa B(NF-κB).Methods:NF-κB protein was extracted and its concentration was determined.DNA probe was labeled by ()~(32)P.The probe was conjugated with NF-κB protein together and underwent electrophoresis.The X-ray film was exposed and autoradiography was obtained.Results:By using this method,highly distinct bands and clear background in the picture were obtained and the activity of NF-κB could be detected by eyes.Conclusion:The modified EMSA is more simple,feasible,and highly sensitive method for detection of DNA-binding proteins.

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Available abstract

Objective:To establish a fast and simple method of electrophoretic mobility shift assay(EMSA)for the identification of DNA-binding protein nuclear factor-kappa B(NF-κB).Methods:NF-κB protein was extracted and its concentration was determined.DNA probe was labeled by ()~(32)P.The probe was conjugated with NF-κB protein together and underwent electrophoresis.The X-ray film was exposed and autoradiography was obtained.Results:By using this method,highly distinct bands and clear background in the picture were obtained and the activity of NF-κB could be detected by eyes.Conclusion:The modified EMSA is more simple,feasible,and highly sensitive method for detection of DNA-binding proteins.

Key concepts: Electrophoretic mobility shift assay, Electrophoresis, DNA, DNA-binding protein, Chemistry, Molecular biology, Nuclear protein, Chromatography

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