Twist-off purification of hair bundles

Methods Mol Biol. 2009;493:241-55. doi: 10.1007/978-1-59745-523-7_14.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dissection / methods*
  • Hair Cells, Auditory / cytology
  • Hair Cells, Auditory / metabolism*
  • Immunoblotting
  • Immunohistochemistry
  • Mass Spectrometry
  • Vestibule, Labyrinth / cytology
  • Vestibule, Labyrinth / metabolism*