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Microwave-Assisted Protein Staining, Destaining, and In-Gel/In-Solution Digestion of Proteins

  • Jennie R. Lill
  • Victor J. Nesatyy
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1853)

Abstract

Rapid evolution of state-of-the-art proteomic analyses has encompassed development of high-throughput analytical instrumentation and bioinformatic tools. However, recently there has been a particular emphasis on increasing the throughput of sample preparation, which has become one of the rate-limiting steps in protein characterization workflows. Researchers have been investigating alternative methods to conventional convection oven incubations to try and reduce sample preparation time for protein characterization. Several protocols have appeared in the literature, which employ microwave irradiation as a tool for the preparation of biological samples for subsequent characterization by a variety of analytical techniques. In this chapter, techniques for microwave-assisted protein staining, destaining, and digestion are described. In general, the application of microwave-assisted technologies resulted in the drastic reduction of overall sample preparation time, though discrepancies in the reproducibility of several published digestion protocols still remain to be clarified.

Key words

Microwave SDS gels Protein staining Destaining Digestion 

Notes

Acknowledgment

Victor J. Nesatyy is grateful to Prof. Yury Tsybin for his support, encouragement, and creative atmosphere while preparing the manuscript.

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Copyright information

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

Authors and Affiliations

  • Jennie R. Lill
    • 1
  • Victor J. Nesatyy
    • 2
    • 3
  1. 1.Department of Protein ChemistryGenentech Inc.South San FranciscoUSA
  2. 2.Biomolecular Mass Spectrometry LaboratoryEPFL-FSB-ISIC LSMBLausanneSwitzerland
  3. 3.Bio21 InstituteUniversity of MelbourneParkvilleAustralia

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