A1 Refereed original research article in a scientific journal
On-target ultrasonic digestion of proteins
Authors: Santos HM, Kouvonen P, Capelo JL, Corthals GL
Publisher: WILEY-BLACKWELL
Publication year: 2013
Journal: Proteomics
Journal name in source: PROTEOMICS
Journal acronym: PROTEOMICS
Number in series: 9
Volume: 13
Issue: 9
First page : 1423
Last page: 1427
Number of pages: 5
ISSN: 1615-9853
DOI: https://doi.org/10.1002/pmic.201200241
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 L followed by 5 min of ultrasonication after trypsin addition. Controls (5 min, 37 degrees 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.
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 L followed by 5 min of ultrasonication after trypsin addition. Controls (5 min, 37 degrees 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.
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