On‐target ultrasonic digestion of proteins |
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| Authors: | Hugo M Santos Petri Kouvonen Jose‐Luis Capelo Garry L Corthals |
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| Institution: | 1. Departamento de Química, Centro de Química Fina e Biotecnologia, (CQFB), REQUIMTE, Faculdade de Ciências e Tecnologia, (FCT), Universidade Nova de Lisboa, (UNL), , Caparica, Portugal;2. Turku Centre for Biotechnology, University of Turku and ?bo Akademi University, , Turku, Finland;3. Department of Biology, Institute of Molecular Systems Biology, ETH, , Zurich, Switzerland |
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| 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. |
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| Keywords: | Accelerated digestion method High throughput On‐target digestion Tissue digestion Ultrasonic digestion |
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