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2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
DOI:

08:23 min

August 06, 2018

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Chapters

  • 00:04Title
  • 00:47Preparation of Plant Cell Material
  • 01:55Tandem Affinity Purification (TAP)
  • 04:59Metabolite and Protein Extraction
  • 06:15Results: Simultaneous Characterization of Protein-Protein and Protein-Metabolite Interactions Employing a Single-Step Tandem Affinity Purification Coupled with Mass Spectrometry
  • 07:23Conclusion

Summary

Automatic Translation

Protein-protein and protein-metabolite interactions are crucial for all cellular functions. Herein, we describe a protocol that allows parallel analysis of these interactions with a protein of choice. Our protocol was optimized for plant cell cultures and combines affinity purification with mass spectrometry-based protein and metabolite detection.

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