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.
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Acknowledgment
Supported by NIH grants R21 DC006097, R01 DC002368, and R01 DC007602 (to P.G.G.), and a DRF grant to J.B.S.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Shin, JB., Pagana, J., Gillespie, P.G. (2009). Twist-Off Purification of Hair Bundles. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology™, vol 493. Humana Press. https://doi.org/10.1007/978-1-59745-523-7_14
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DOI: https://doi.org/10.1007/978-1-59745-523-7_14
Publisher Name: Humana Press
Print ISBN: 978-1-934115-62-6
Online ISBN: 978-1-59745-523-7
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