F. Fischer and B. Lorber contributed equally to this work.
Protein analysis by dynamic light scattering: Methods and techniques for students†
Article first published online: 10 OCT 2012
Copyright © 2012 Wiley Periodicals, Inc.
Biochemistry and Molecular Biology Education
Volume 40, Issue 6, pages 372–382, November/December 2012
How to Cite
Lorber, B., Fischer, F., Bailly, M., Roy, H. and Kern, D. (2012), Protein analysis by dynamic light scattering: Methods and techniques for students. Biochem. Mol. Biol. Educ., 40: 372–382. doi: 10.1002/bmb.20644
This work is supported by the Université de Strasbourg, the French Ministère de l'Enseignement Supérieur et de la Recherche, the Association pour la Recherche sur le Cancer (ARC), the Centre National de la Recherche Scientifique (CNRS), French National Program Investissement d'Avenir (Labex Mitocross).
- Issue published online: 19 NOV 2012
- Article first published online: 10 OCT 2012
- Manuscript Accepted: 23 JUL 2012
- Manuscript Received: 23 MAY 2012
- Université de Strasbourg, the French Ministère de l'Enseignement Supérieur et de la Recherche, the Association pour la Recherche sur le Cancer
- the Centre National de la Recherche Scientifique (CNRS)
- French National Program Investissement d'Avenir (Labex Mitocross)
- Light scattering;
Dynamic light scattering (DLS) analyses are routinely used in biology laboratories to detect aggregates in macromolecular solutions, to determine the size of proteins, nucleic acids, and complexes or to monitor the binding of ligands. This article is written for graduate and undergraduate students with access to DLS and for faculty members who wish to incorporate DLS into a lab activity, a practical course or research. It reviews the basic concepts of light scattering measurements and addresses four critical aspects of the analysis and interpretation of DLS results. To ensure reproducible quantitative data, attention should be paid to controlling the preparation and handling of proteins or assemblies because variations in the state of aggregation, induced by minor changes in experimental condition or technique, might compromise DLS results and affect protein activity. Variables like temperature, solvent viscosity, and inter-particle interactions may also influence particle size determination. Every point is illustrated by case studies, including a commercially available albumin, a small RNA virus isolated from plants, as well as four soluble proteins and a ribonucleoprotein assembly purified and characterized by students in the frame of their master degree. © 2012 by The International Union of Biochemistry and Molecular Biology