Auditory and Vestibular Research pp 241-255

Part of the Methods in Molecular Biology™ book series (MIMB, volume 493) | Cite as

Twist-Off Purification of Hair Bundles

  • Jung-Bum Shin
  • James Pagana
  • Peter G. Gillespie

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.

Keywords

Auditory vestibular hair bundles mechanotransduction agarose mass spectrometry 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Jung-Bum Shin
    • 1
  • James Pagana
    • 1
  • Peter G. Gillespie
    • 1
  1. 1.Oregon Hearing Research Center and Vollum InstituteOregon Health&Science UniversityPortlandUSA

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