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JoVE Journal
Biochemistry

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Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
 

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Article DOI: 10.3791/60432-v 11:19 min November 17th, 2019
November 17th, 2019

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résumé

Described is a proteomics workflow for identifying protein interaction partners from a nuclear subcellular fraction using immunoaffinity enrichment of a given protein of interest and label-free mass spectrometry. The workflow includes subcellular fractionation, immunoprecipitation, filter aided sample preparation, offline cleanup, mass spectrometry, and a downstream bioinformatics pipeline.

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Keywords: Label-free Immunoprecipitation Mass Spectrometry Nuclear Interactome Protein-protein Interactions Subcellular Fractionation Affinity Purification Proteomics Workflow Cell Lysis Nuclear Lysate Dounce Homogenization
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