Journal
/
Biology
/
Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures

Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures

DOI:
10.3791/50535-v

•

11:44 min

•

September 28, 2013

•

, ,
1Signaling Proteomics,Max Plank Institute of Molecular Plant Physiology, 2Department of Plant Systems Biology,University of Hohenheim

Chapters

  • 00:05Title
  • 01:35Plasma Membrane Purification
  • 03:56Detergent Resistant Membrane Preparation
  • 05:33Extraction of Proteins from the Detergent Resistant Fraction and In-solution Trypsin Digestion
  • 07:04Use of C18-StageTips for Peptide Desalting and Concentration
  • 08:48Results: Comparative Proteomic Analysis
  • 11:00Conclusion

Summary

Automatic Translation

Automatic Translation

Here we describe a robust method for the fractionation of plant plasma membranes into detergent resistant and detergent soluble membranes based on a mixture of unlabeled and in vivo fully 15N labeled Arabidopsis thaliana cell cultures. The procedure is applied for comparative proteomic studies to understand signaling processes.

Tags

Metabolic LabelingMembrane FractionationComparative ProteomicsArabidopsis ThalianaSuspension Cell CulturesPlasma MembraneMicrodomainsDetergent-resistant Membrane FractionSucrose GradientQuantitative ProteomicsK15NO3 Labeling

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

Toxin Induction and Protein Extraction from Fusarium spp. Cultures for Proteomic Studies

Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::β-glucuronidase Reporter Gene Expression

Measuring Cell Cycle Progression Kinetics with Metabolic Labeling and Flow Cytometry

A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass Spectrometry

Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements

Real Time Analysis of Metabolic Profile in Ex Vivo Mouse Intestinal Crypt Organoid Cultures

Mixed Primary Cultures of Murine Small Intestine Intended for the Study of Gut Hormone Secretion and Live Cell Imaging of Enteroendocrine Cells

Analysis of Arabidopsis thaliana Growth Behavior in Different Light Qualities

Author Spotlight: Image-Based Methods to Study Membrane Trafficking Events in Stomatal Lineage Cells

Read Article