JoVE Logo
Faculty Resource Center

Sign In

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

674 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

-- Views

Related Videos

article

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

article

High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution

article

Author Spotlight: Characterizing DNA G-Quadruplex by Bis-3-Chloropiperidine Based Chemical Mapping

article

Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals

article

Affinity Purification of a Fibrinolytic Enzyme from Sipunculus nudus

article

Author Spotlight: Peptidome Extraction from Small Extracellular Vesicles Isolated from Bone Marrow-Derived Macrophages

article

Author Spotlight: A Novel Method for Comprehensive Cell Component Analysis of Cerebral Blood Clots

article

Author Spotlight: Efficient Venom Extraction Method from Trichogramma Parasitoid Wasps

article

Author Spotlight: Advancing EVtrap for High-Throughput Proteomics in Disease Biomarker Discovery

article

Author Spotlight: Enhancing CryoEM Sample Preparation Using Graphene Monolayer on Microscopy Grids

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved