On-target ultrasonic digestion of proteins

H. M. Santos, P. Kouvonen, J. L. Capelo, G. L. Corthals

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

In the present work, we report a novel on-target protein cleavage method. The method utilizes ultrasonic energy and allows up to 20 samples to be cleaved in 5 min for protein identification and one sample in 30 s for on-tissue digestion. The standard proteins were spotted on a conductive glass slide in a volume of 0.5 μL followed by 5 min of ultrasonication after trypsin addition. Controls (5 min, 37°C no ultrasonication) were also assayed. After trypsin addition, digestion of the tissues was enhanced by 30 s of ultrasonication. The samples were analyzed and compared to those obtained by using conventional 3 h heating proteolysis. The low sample volume needed for the digestion and reduction in sample-handling steps and time are the features that make this method appealing to the many laboratories working with high-throughput sample treatment. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Original languageEnglish
Pages (from-to)1423-1427
Number of pages5
JournalProteomics
Volume13
Issue number9
DOIs
Publication statusPublished - May 2013

Keywords

  • Accelerated digestion method
  • High throughput
  • On-target digestion
  • Tissue digestion
  • Ultrasonic digestion
  • protein
  • article
  • controlled study
  • heating
  • nonhuman
  • priority journal
  • protein analysis
  • protein degradation
  • proteomics
  • reduction
  • ultrasound
  • Animals
  • Equipment Design
  • Liver
  • Mice
  • Proteins
  • Proteomics
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Trypsin
  • Ultrasonics

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