Rivista
/
Biologia
/
Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures
JoVE Journal
Biologia
È necessario avere un abbonamento a JoVE per visualizzare questo.  Accedi o inizia la tua prova gratuita.
JoVE Journal Biologia
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

Capitoli

  • 00:05Titolo
  • 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

Traduzione automatica

Traduzione automatica

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

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

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

check_url/it/50535?article_type=v

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay
check_url/it/50535?article_type=v

Toxin Induction and Protein Extraction from Fusarium spp. Cultures for Proteomic Studies
check_url/it/50535?article_type=v

Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::β-glucuronidase Reporter Gene Expression
check_url/it/50535?article_type=v

Measuring Cell Cycle Progression Kinetics with Metabolic Labeling and Flow Cytometry
check_url/it/50535?article_type=v

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate
check_url/it/50535?article_type=v

A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass Spectrometry
check_url/it/50535?article_type=v

Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements
check_url/it/50535?article_type=v

Real Time Analysis of Metabolic Profile in Ex Vivo Mouse Intestinal Crypt Organoid Cultures
check_url/it/50535?article_type=v

Mixed Primary Cultures of Murine Small Intestine Intended for the Study of Gut Hormone Secretion and Live Cell Imaging of Enteroendocrine Cells
check_url/it/50535?article_type=v

Analysis of Arabidopsis thaliana Growth Behavior in Different Light Qualities

Read Article