Protocol

Cytoskeleton Methods and Protocols

Volume 586 of the series Methods in Molecular Biology pp 207-225

Date:

Purification and Localization of Intraflagellar Transport Particles and Polypeptides

  • Roger D. SlobodaAffiliated withDepartment of Biological Sciences, Dartmouth College

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Summary

The growth and maintenance of almost all cilia and flagella are dependent on the proper functioning of the process of intraflagellar transport (IFT). This includes the primary cilia of most cells in humans that are in interphase or the Go phase of the cell cycle. The model system for the study of IFT is the flagella of the bi-flagellate green alga Chlamydomonas. It is in this organism that IFT was first discovered, and genetic data from a Chlamydomonas mutant first linked the process of IFT to polycystic kidney disease in humans. The information given in this chapter addresses procedures to purify IFT particles from flagella and localize these particles, and their associated motor proteins, in flagella using light and electron microscopic approaches.

Key words:

Flagella Cilia Intraflagellar transport Motility Immunofluorescence Immunogold EM