Twist-off purification of hair bundles.
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.
Shin
Jung-Bum
JB
Oregon Hearing Research Center and Vollum Institute, Oregon Health & Science University, Portland, OR, USA.
Pagana
James
J
Gillespie
Peter G
PG
D004210:Dissection / Q000379:methods*; D006198:Hair Cells, Auditory / Q000166:cytology / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D013058:Mass Spectrometry; D014722:Vestibule, Labyrinth / Q000166:cytology / Q000378:metabolism*