Twist-off purification of hair bundles

Jung Bum Shin, James Pagana, Peter G. Gillespie

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

Abstract

Purification of hair bundles from inner-ear organs allows biochemical analysis of bundle constituents, including proteins and lipids. We describe here the "twist-off" method of bundle isolation, where dissected inner-ear organs are embedded in agarose, then subjected to a mechanical disruption that shears off bundles and leaves them in agarose blocks. With care in the dissection and in clean-up of the isolated bundles, contamination from cell bodies can be kept to a minimum. Isolated bundles can be analyzed by a variety of techniques, including immunocytochemistry, SDS-PAGE, immunoblotting, and mass spectrometry.

Original languageEnglish (US)
Title of host publicationAuditory and Vestibular Research
Subtitle of host publicationMethods and Protocols
PublisherHumana Press
Pages241-255
Number of pages15
ISBN (Print)9781934115626
DOIs
StatePublished - 2009

Publication series

NameMethods in Molecular Biology
Volume493
ISSN (Print)1064-3745

Keywords

  • Agarose
  • Auditory
  • Hair bundles
  • Mass spectrometry
  • Mechanotransduction
  • Vestibular

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Shin, J. B., Pagana, J., & Gillespie, P. G. (2009). Twist-off purification of hair bundles. In Auditory and Vestibular Research: Methods and Protocols (pp. 241-255). (Methods in Molecular Biology; Vol. 493). Humana Press. https://doi.org/10.1007/978-1-59745-523-7_14