title: On-target ultrasonic digestion of proteins.
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.
authors: Departamento 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
authors: Hugo M
authors: HM
authors: Santos
authors: Petri
authors: P
authors: Kouvonen
authors: Jose-Luis
authors: JL
authors: Capelo
authors: Garry L
authors: GL
authors: Corthals
mesh_terms: D000818:Animals; D004867:Equipment Design; D008099:Liver / Q000378:metabolism; D051379:Mice; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry; D014465:Ultrasonics / Q000295:instrumentation / Q000379:methods*
chemical_list: D011506:Proteins; D014357:Trypsin