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Immunoprecipitation coupled with mass spectrometry (IP-MS) is a powerful method that enables the identification of protein–protein interactions and isolation of protein complexes. When optimized to work with plant material, it proved to be suitable for the isolation of different types of nuclear complexes from Arabidopsis. Here, we describe a detailed protocol for the isolation of chromatin remodeling and polycomb repressive complexes using Arabidopsis lines that express GFP-tagged proteins as baits, directly from whole-cell extracts or after nuclei enrichment. We describe different experimental variations of the key steps in the protocol and discuss the analysis and interpretation of the IP-MS data.

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Isolation and Identification of Nuclear Protein Complexes Using GFP-Tagged Arabidopsis Lines and IP-MS Approach

  • 实验方案

  • 发表日期：

    2025年

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  • Paulina Stachula,
  • Rafał Archacki

摘要

Immunoprecipitation coupled with mass spectrometry (IP-MS) is a powerful method that enables the identification of protein–protein interactions and isolation of protein complexes. When optimized to work with plant material, it proved to be suitable for the isolation of different types of nuclear complexes from Arabidopsis. Here, we describe a detailed protocol for the isolation of chromatin remodeling and polycomb repressive complexes using Arabidopsis lines that express GFP-tagged proteins as baits, directly from whole-cell extracts or after nuclei enrichment. We describe different experimental variations of the key steps in the protocol and discuss the analysis and interpretation of the IP-MS data.