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Preparation of Cell Lysates of Fission Yeast for Immunoprecipitation

  • Akihisa MatsuyamaEmail author
  • Atsuko Shirai
  • Minoru Yoshida
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1721)

Abstract

Immunoprecipitation is one of the most important and widely used techniques for the detection and purification of a protein of interest. Thanks to highly specific interaction between antigen and antibody, a target protein is purified and concentrated effectively. To obtain reasonable amounts of a target protein, it is crucially important to prepare total cell lysates in which the target protein is present in a soluble form. Here, we describe methods to prepare total cell lysates of fission yeast, which are then used directly for immunoprecipitation. We also describe some tips to select reagents for preparing buffers having a substantial impact on protein solubility, because there is essentially no reagent that can accommodate the full range of proteins having different characteristics.

Key words

Total cell lysate Immunoprecipitation Solubilization 

Notes

Acknowledgments

This work was supported by JSPS KAKENHI Grant Number 15K14925.

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Akihisa Matsuyama
    • 1
    • 2
    Email author
  • Atsuko Shirai
    • 2
    • 3
  • Minoru Yoshida
    • 1
    • 2
    • 4
  1. 1.Chemical Genomics Research GroupRIKEN Center for Sustainable Resource ScienceSaitamaJapan
  2. 2.Chemical Genetics Laboratory, RIKENSaitamaJapan
  3. 3.Cellular Memory Laboratory, RIKENSaitamaJapan
  4. 4.Department of Biotechnology,The University of TokyoJapan

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