Protocol

Cytoskeleton Methods and Protocols

Volume 1365 of the series Methods in Molecular Biology pp 119-137

Purification and Localization of Intraflagellar Transport Particles and Polypeptides

  • Roger D. SlobodaAffiliated withDepartment of Biological Sciences, Dartmouth CollegeThe Marine Biological Laboratory Email author 

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Abstract

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 human cells that are in the Go phase of the cell cycle. The model system for the study of IFT is the flagella of the biflagellate 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 provided 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 IFT Motility Immunofluorescence Immunogold EM