JoVE Logo
Centre de ressources universitaires

S'identifier

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

DOI :

10.3791/58754-v

January 7th, 2019

January 7th, 2019

9,177 Views

1Van Andel Institute, 2Vollum Institute, 3Janelia Research Campus

This protocol describes techniques used to determine ion channel structures by cryo-electron microscopy, including a baculovirus system used to efficiently express genes in mammalian cells with minimum effort and toxicity, protein extraction, purification, and quality checking, sample grid preparation and screening, as well as data collection and processing.

Tags

Lipid sensitive Cation Channel TRPC3

-- Vues

Vidéos Connexes

article

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

article

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

article

Single Particle Cryo-Electron Microscopy: From Sample to Structure

article

Manual Blot-and-Plunge Freezing of Biological Specimens for Single-Particle Cryogenic Electron Microscopy

article

Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows

article

A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion

article

Preparation of Nucleosome Core Particles Complexed with DNA Repair Factors for Cryo-Electron Microscopy Structural Determination

article

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon

article

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

article

High-Throughput Expression and Purification of Human Solute Carriers for Structural and Biochemical Studies

JoVE Logo

Confidentialité

Conditions d'utilisation

Politiques

Recherche

Enseignement

À PROPOS DE JoVE

Copyright © 2024 MyJoVE Corporation. Tous droits réservés.