Measurement and analysis of in vitro actin polymerization

Lynda K. Doolittle, Michael K. Rosen, Shae B. Padrick

Research output: Chapter in Book/Report/Conference proceedingChapter

28 Scopus citations

Abstract

The polymerization of actin underlies force generation in numerous cellular processes. While actin polymerization can occur spontaneously, cells maintain control over this important process by preventing actin filament nucleation and then allowing stimulated polymerization and elongation by several regulated factors. Actin polymerization, regulated nucleation, and controlled elongation activities can be reconstituted in vitro, and used to probe the signaling cascades cells use to control when and where actin polymerization occurs. Introducing a pyrene fluorophore allows detection of filament formation by an increase in pyrene fluorescence. This method has been used for many years and continues to be broadly used, owing to its simplicity and flexibility. Here we describe how to perform and analyze these in vitro actin polymerization assays, with an emphasis on extracting useful descriptive parameters from kinetic data.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages273-293
Number of pages21
Volume1046
ISBN (Print)9781627035378
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1046
ISSN (Print)10643745

Keywords

  • Actin polymerization
  • Biochemical assay
  • In vitro reconstitution
  • Quantitative analysis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Doolittle, L. K., Rosen, M. K., & Padrick, S. B. (2013). Measurement and analysis of in vitro actin polymerization. In Methods in Molecular Biology (Vol. 1046, pp. 273-293). (Methods in Molecular Biology; Vol. 1046). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-538-5_16