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JoVE Journal
Biochemistry

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Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry
 

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Article DOI: 10.3791/59865-v 08:07 min July 26th, 2019
July 26th, 2019

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Summary

This protocol focuses on the identification of proteins that bind to inositol phosphates or phosphoinositides. It uses affinity chromatography with biotinylated inositol phosphates or phosphoinositides that are immobilized via streptavidin to agarose or magnetic beads. Inositol phosphate or phosphoinositide binding proteins are identified by Western blotting or mass spectrometry.

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Keywords: Inositol Phosphate Phosphoinositide Protein Binding Affinity Chromatography Western Blot Mass Spectrometry Regulatory Function Signal Transduction Cell Development Biotinylated Lyposome Free T. Brucei PBS-G Lysis Buffer Protease Inhibitor Phosphatase Inhibitor Binding Assay
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