Title

Protein Analysis By Dynamic Light Scattering: Methods And Techniques For Students

Keywords

Light scattering; Protein; Temperature; Viscosity

Abstract

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 Wiley Periodicals, Inc.

Publication Date

11-1-2012

Publication Title

Biochemistry and Molecular Biology Education

Volume

40

Issue

6

Number of Pages

372-382

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1002/bmb.20644

Socpus ID

84869789038 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/84869789038

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