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A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis
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Bioquímica
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JoVE Journal Bioquímica
A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis

A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis

DOI:
10.3791/55077-v

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05:43 min

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January 24, 2017

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, , , ,
1Graduate School of Frontier Biosciences,Osaka University, 2Department of Biotechnology and Genetic Engineering,Jahangirnagar University, 3Division of Cell Regeneration and Transplantation,School of Medicine, Nihon University, 4Institute of Genetics and Animal Breeding of the Polish Academy of Sciences, 5Graduate School of Life and Medical Sciences,Doshisha University

Capítulos

  • 00:05Título
  • 00:45Protein Extraction
  • 02:01Immunoprecipitation
  • 03:47Alkylation and Trypsinization
  • 04:46Results: Purification of ARIP4 Complexes
  • 05:13Conclusion

Summary

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We have established a method for the purification of coregulatory interaction proteins using the LC-MS/MS system.

Tags

Protein PreparationHigh-throughputProtein InteractionsNuclear CofactorLC-MS/MS AnalysisTranscriptional MechanismsGene RegulationFlag-tagged ARIP4HEK 293 CellsCell HarvestingCell LysisImmunoprecipitationProtein ElutionMass Spectrometry

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