Fluorescence-based proteasome activity profiling

Jong, A., Schuurman, K.G., Rodenko, B., Ovaa, H. and Berkers, C.R. (2012) Fluorescence-based proteasome activity profiling. Methods in Molecular Biology, 803, pp. 183-204. (doi: 10.1007/978-1-61779-364-6_13)

Abstract

With the proteasome emerging as a therapeutic target for cancer treatment, accurate tools for monitoring proteasome (inhibitor) activity are in demand. In this chapter, we describe the synthesis and use of a fluorescent proteasome activity probe that allows for accurate profiling of proteasomal activity in cell lysates, intact cells, and murine and human patient-derived material, with high sensitivity using SDS-PAGE. The probe allows for direct scanning of the gel for fluorescent emission of the distinct proteasomal subunits and circumvents the use of Western blot analysis. Due to its suitable biochemical and biophysical properties, the fluorescent probe can also be used for confocal laser scanning microscopy and flow cytometry-based experiments.

• Item Type: Articles
• Additional Information: The original publication is available at www.springerlink.com
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Rodenko, Dr Boris and Berkers, Dr Celia
• Authors: Jong, A., Schuurman, K.G., Rodenko, B., Ovaa, H., and Berkers, C.R.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Methods in Molecular Biology
• ISSN: 1064-3745
• ISSN (Online): 1940-6029
• Published Online: 01 January 2012
• Copyright Holders: Copyright © 2012 Springer Science+Business Media, LLC.
• First Published: First published in Methods in Molecular Biology 803:183-204
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher