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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.

Transcript

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Inositol Phosphate Phosphoinositide Affinity Chromatography Western Blot Mass Spectrometry Regulatory Functions Gene Expression Protein Trafficking Signal Transduction Cell Development Protein Binding Conformational Change Catalytic Activity Protein Interactions Lyposome Free Commercially Available Reagents Biotinylated Inositol Phosphate Blood Stream Forms T.brucei Cells HMI-9 Media FBS Supplementation Cell Density
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