Applied Biochemistry and Biotechnology

, Volume 163, Issue 1, pp 64–70 | Cite as

Use of Highly Purified and Mixed Antibodies for Simultaneous Detection of Multiple Protein Species Released from Mitochondria upon Induction of the Permeability Transition

  • Takenori Yamamoto
  • Mizuki Ohashi
  • Sho Mizutani
  • Yuuya Yoshimura
  • Eriko Obana
  • Hiroshi Terada
  • Yasuo ShinoharaEmail author


Concomitant with the induction of the mitochondrial permeability transition (PT), cytochrome c is released from mitochondria into the cytosol where it triggers subsequent steps of cellular apoptosis. Thus, inducers of the mitochondrial PT would become “seed compounds” of regulators of apoptosis. However, when we examine the actions of certain chemicals on the release of mitochondrial cytochrome c, the behaviors of not only cytochrome c but also multiple mitochondrial protein species must be carefully examined because the mitochondrial PT and release of proteins from mitochondria occur in diverse manners. In the present study, we examined whether it is possible to measure the behaviors of multiple protein species in a single experiment using purified and mixed antibodies. The results obtained clearly indicate that this procedure would be applicable for high-throughput screening of regulators of apoptosis. Further requirements necessary for the establishment of a useful screening system for apoptosis regulators are discussed.


Mitochondria Permeability transition Protein release Cytochrome c Western blotting 


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Takenori Yamamoto
    • 1
  • Mizuki Ohashi
    • 1
    • 2
  • Sho Mizutani
    • 1
    • 2
  • Yuuya Yoshimura
    • 1
    • 2
  • Eriko Obana
    • 1
    • 2
  • Hiroshi Terada
    • 3
  • Yasuo Shinohara
    • 1
    • 2
    Email author
  1. 1.Institute for Genome ResearchUniversity of TokushimaTokushimaJapan
  2. 2.Faculty of Pharmaceutical SciencesUniversity of TokushimaTokushimaJapan
  3. 3.Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan

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