Journal
/
Bioquímica
/
Nitrogen Cavitation and Differential Centrifugation Allows for Monitoring the Distribution of Peripheral Membrane Proteins in Cultured Cells
JoVE Revista
Bioquímica
Se requiere una suscripción a JoVE para ver este contenido.  Inicie sesión o comience su prueba gratuita.
JoVE Revista Bioquímica
Nitrogen Cavitation and Differential Centrifugation Allows for Monitoring the Distribution of Peripheral Membrane Proteins in Cultured Cells

Nitrogen Cavitation and Differential Centrifugation Allows for Monitoring the Distribution of Peripheral Membrane Proteins in Cultured Cells

DOI:
10.3791/56037-v

•

08:24 min

•

August 18, 2017

•

,
1Langone Medical Center,New York University

Capítulos

  • 00:05Título
  • 00:41Cell Harvesting
  • 01:35Nitrogen Cavitation
  • 03:31Separation of Cytosolic and Membrane Fractions
  • 05:22Results: Partitioning of Cellular Proteins in Membrane and Cytosolic Fractions
  • 07:07Conclusion

Summary

Traducción Automática

Traducción Automática

Here we present protocols for detergent-free homogenization of cultured mammalian cells based on nitrogen cavitation and subsequent separation of cytosolic and membrane-bound proteins by ultracentrifugation. This method is ideal for monitoring the partitioning of peripheral membrane proteins between soluble and membrane fractions.

Tags

Nitrogen CavitationDifferential CentrifugationPeripheral Membrane ProteinsCultured CellsSubcellular CompartmentalizationHEK293 CellsHomogenization BufferPressurizationProtease InhibitorCell DisruptionFractionationSoluble FractionMembrane Fraction

Article

Embed

Añadir a la lista de reproducción

Usage Statistics

Videos relacionados

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

Preparation of Giant Vesicles Encapsulating Microspheres by Centrifugation of a Water-in-oil Emulsion

Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells

Analyses of Mitochondrial Calcium Influx in Isolated Mitochondria and Cultured Cells

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

Transverse Sectioning of Mature Rice (Oryza sativa L.) Kernels for Scanning Electron Microscopy Imaging Using Pipette Tips as Immobilization Support

Cell-Free Production of Proteoliposomes for Functional Analysis and Antibody Development Targeting Membrane Proteins

Read Article