Unit

UNIT 13.2 In Vitro Motility Assay to Study Translocation of Actin by Myosin

  1. James R. Sellers

Published Online: 1 MAY 2001

DOI: 10.1002/0471143030.cb1302s00

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Sellers, J. R. 2001. In Vitro Motility Assay to Study Translocation of Actin by Myosin. Current Protocols in Cell Biology. 00:13.2:13.2.1–13.2.10.

Author Information

  1. National Heart, Lung, and Blood Institute, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1998

Abstract

A basic property of myosin is its ability to interact with and translocate actin. This unit describes an in vitro motility assay that can be used to study the translocation, or sliding, of actin filaments by myosin bound to a coverslip. The assay makes use of the ability to image single F-actin filaments labeled with rhodamine phalloidin, a high-affinity fluorescent ligand using fluorescence microscopy. The system is fast, easy to set up and maintain, uses only small amounts of protein, and yields quantitative results.