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 language | English |
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Pages (from-to) | 1423-1427 |
Number of pages | 5 |
Journal | Proteomics |
Volume | 13 |
Issue number | 9 |
DOIs | |
Publication status | Published - 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