Detection and Identification of Low-Abundant Proteins Using HPE Gels, Fluorescent Stains, and MALDI-ToF-ToF-MS

  • Martin Moche
  • Dirk Albrecht
  • Reiner Westermeier
  • Knut BüttnerEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1841)


Two-dimensional electrophoresis as a complementary approach to gel-free proteomic methods possesses the ability to separate physiologically important isoforms of proteins in an unbiased manner. Frequently, those isoforms are low-abundant regulators, and therefore, detection and identification of low-abundant proteins is highly necessary to exploit this advantage. We describe an experimental sequence of classical operations to process gels but optimized them, in order to identify each detectable protein spot on gel.

Key words

High performance electrophoresis Low-abundant proteins Fluorescent stains Scanning conditions Tryptic digestion MALDI-ToF-MS 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Martin Moche
    • 1
  • Dirk Albrecht
    • 2
  • Reiner Westermeier
    • 3
  • Knut Büttner
    • 2
    Email author
  1. 1.Waters GmbHEschbornGermany
  2. 2.Department of Microbial Physiology and Molecular Biology, Institute for MicrobiologyUniversity GreifswaldGreifswaldGermany
  3. 3.FreisingGermany

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