A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
On-target ultrasonic digestion of proteins
Tekijät: Santos HM, Kouvonen P, Capelo JL, Corthals GL
Kustantaja: WILEY-BLACKWELL
Julkaisuvuosi: 2013
Lehti: Proteomics
Tietokannassa oleva lehden nimi: PROTEOMICS
Lehden akronyymi: PROTEOMICS
Numero sarjassa: 9
Vuosikerta: 13
Numero: 9
Aloitussivu: 1423
Lopetussivu: 1427
Sivujen määrä: 5
ISSN: 1615-9853
DOI: https://doi.org/10.1002/pmic.201200241
Tiivistelmä
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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