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Updated: May 13, 2026

Evaluation of Protein–Protein Interactions using an On-Membrane Digestion Technique
Published on: July 19, 2019
On-target ultrasonic digestion of proteins
Hugo M Santos1, Petri Kouvonen, Jose-Luis Capelo
1Departamento de Química, Centro de Química Fina e Biotecnologia, (CQFB), REQUIMTE, Faculdade de Ciências e Tecnologia, (FCT), Universidade Nova de Lisboa, (UNL), Caparica, Portugal. hms14862@fct.unl.pt
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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