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Extracting Modified Microtubules from Mammalian Cells to Study Microtubule-Protein Complexes by Cryo-Electron Microscopy

DOI :

10.3791/65126-v

8:02 min

March 3rd, 2023

March 3rd, 2023

750 Views

1Oncode Institute and Division of Biochemistry, Netherlands Cancer Institute

Here, we describe a protocol to extract endogenous tubulin from mammalian cells, which can lack or contain specific microtubule-modifying enzymes, to obtain microtubules enriched for a specific modification. We then describe how the extracted microtubules can be decorated with purified microtubule-binding proteins to prepare grids for cryo-electron microscopy.

Tags

Modified Microtubules

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