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

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11:44 min

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September 28, 2013

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, ,
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

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

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