Abstract: Objective To establish a method of cilia isolation from airway epithelium and provide the methods of further studying the role of the protein phosphorylation in the mechanisms of ciliary motility. Methods The rabbit trachea epithelium was incubated in isolation buffer containing Triton X-100 in order to isolate the cilia from the epithelial tissue. Cilia were then collected by centrifugation. The sample was examined by electron microscopy, and the β-tubulin and the phosphorylated proteins were visualized by Western blotting. Results 1 The electron micrograph demonstrated a highly enriched ciliary preparation showing the characteristic features of intact axoneme; 2 Western blotting analysis showed visible bands of β-tubulin; 3 Western blotting analysis detected five tyrosine phosphorylated bands. Conclusion The ciliary preparation can be obtained by incubating the rabbit airway epithelium in isolation buffer containing Triton X-100 and the following centrifugation. The preparation can be used to study the role of the protein phosphorylation in the mechanisms of ciliary motility.

Key words: cilia, isolation, phosphorylation