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Statements

Subject Item
n2:RIV%2F61989592%3A15410%2F03%3A00001376%21RIV%2F2004%2FMSM%2F154104%2FN
rdf:type
skos:Concept n16:Vysledek
dcterms:description
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.
dcterms:title
Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins
skos:prefLabel
Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins Fast-Response Proteomics by Accelerated In-Gel Digestion of Proteins
skos:notation
RIV/61989592:15410/03:00001376!RIV/2004/MSM/154104/N
n3:strany
1300-1306
n3:aktivita
n15:Z
n3:aktivity
Z(MSM 153100013)
n3:cisloPeriodika
6
n3:dodaniDat
n10:2004
n3:domaciTvurceVysledku
n12:9708790
n3:druhVysledku
n17:J
n3:duvernostUdaju
n14:S
n3:entitaPredkladatele
n5:predkladatel
n3:idSjednocenehoVysledku
607106
n3:idVysledku
RIV/61989592:15410/03:00001376
n3:jazykVysledku
n7:eng
n3:klicovaSlova
FLIGHT MASS-SPECTROMETRY, FUNCTIONAL PROTEOMICS, POLYACRYLAMIDE GELS, TRYPTIC DIGESTION, IDENTIFICATION, QUADRUPOLE, TOOL, COMPLEXES, PEPTIDES, TRYPSIN
n3:klicoveSlovo
n4:QUADRUPOLE n4:TRYPTIC%20DIGESTION n4:TRYPSIN n4:POLYACRYLAMIDE%20GELS n4:IDENTIFICATION n4:PEPTIDES n4:TOOL n4:COMPLEXES n4:FLIGHT%20MASS-SPECTROMETRY n4:FUNCTIONAL%20PROTEOMICS
n3:kodStatuVydavatele
US - Spojené státy americké
n3:kontrolniKodProRIV
[9A2D065F2A9E]
n3:nazevZdroje
Analytical Chemistry
n3:obor
n18:CE
n3:pocetDomacichTvurcuVysledku
1
n3:pocetTvurcuVysledku
4
n3:pocetUcastnikuAkce
0
n3:pocetZahranicnichUcastnikuAkce
0
n3:rokUplatneniVysledku
n10:2003
n3:svazekPeriodika
75
n3:tvurceVysledku
Šebela, Marek Havliš, Jan Thomas, Henrik Shevchenko, Andrej
n3:zamer
n6:MSM%20153100013
s:issn
0003-2700
s:numberOfPages
7
n8:organizacniJednotka
15410