About: Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins     Goto   Sponge   NotDistinct   Permalink

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  • Kinetics of in-gel digestion of proteins by modified and native trypsins was studied by MALDI TOF mass spectrometry using O-18-labeled peptides as internal standards. The effect of the temperature, enzyme concentration, digestion time, and surface area of gel pieces on the yield of digestion products was characterized. Based on the kinetic data, we developed a protocol that enabled the identification of gel-separated proteins with 30-min digestion time without compromising the peptide yield and the sensitivity compared to conventional protocols that typically rely upon overnight enzymatic cleavage. The accelerated digestion protocol was tested in identification of more than 120 proteins from budding and fission yeasts at the subpicomole level.
  • Kinetics of in-gel digestion of proteins by modified and native trypsins was studied by MALDI TOF mass spectrometry using O-18-labeled peptides as internal standards. The effect of the temperature, enzyme concentration, digestion time, and surface area of gel pieces on the yield of digestion products was characterized. Based on the kinetic data, we developed a protocol that enabled the identification of gel-separated proteins with 30-min digestion time without compromising the peptide yield and the sensitivity compared to conventional protocols that typically rely upon overnight enzymatic cleavage. The accelerated digestion protocol was tested in identification of more than 120 proteins from budding and fission yeasts at the subpicomole level. (en)
Title
  • Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins
  • Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins (en)
skos:prefLabel
  • Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins
  • Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins (en)
skos:notation
  • RIV/61989592:15410/03:00001376!RIV/2004/MSM/154104/N
http://linked.open.../vavai/riv/strany
  • 1300-1306
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • Z(MSM 153100013)
http://linked.open...iv/cisloPeriodika
  • 6
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 607106
http://linked.open...ai/riv/idVysledku
  • RIV/61989592:15410/03:00001376
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • FLIGHT MASS-SPECTROMETRY, FUNCTIONAL PROTEOMICS, POLYACRYLAMIDE GELS, TRYPTIC DIGESTION, IDENTIFICATION, QUADRUPOLE, TOOL, COMPLEXES, PEPTIDES, TRYPSIN (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • US - Spojené státy americké
http://linked.open...ontrolniKodProRIV
  • [9A2D065F2A9E]
http://linked.open...i/riv/nazevZdroje
  • Analytical Chemistry
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...ocetUcastnikuAkce
http://linked.open...nichUcastnikuAkce
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 75
http://linked.open...iv/tvurceVysledku
  • Šebela, Marek
  • Havliš, Jan
  • Shevchenko, Andrej
  • Thomas, Henrik
http://linked.open...n/vavai/riv/zamer
issn
  • 0003-2700
number of pages
http://localhost/t...ganizacniJednotka
  • 15410
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