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Eukaryotic flagella are conserved multifunctional organelles with roles in motility, intercellular interactions and signal transduction. Leishmania possess a single flagellum at all stages of their life cycle. Flagella of promastigote forms in the fly are long and motile, with a canonical 9+2 microtubule axoneme and an extra-axonemal paraflagellar rod (PFR). This protocol describes a simple method for the isolation of Leishmania mexicana promastigote flagella, optimised to yield intact flagella that retain both the cytoskeletal elements (9+2 axoneme and PFR) and the surrounding membrane. The isolated flagella and deflagellated cell bodies are suitable for analysis by electron microscopy, protein mass spectrometry and lipidomics.

Type

Journal article

Journal

Methods in Molecular Biology

Publisher

Humana Press