2017
DOI: 10.1038/srep43512
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Evaluation of dynamic changes in interstitial fluid proteome following microdialysis probe insertion trauma in trapezius muscle of healthy women

Abstract: Microdialysis (MD) has been shown to be a promising technique for sampling of biomarkers. Implantation of MD probe causes an acute tissue trauma and provokes innate response cascades. In order to normalize tissue a two hours equilibration period for analysis of small molecules has been reported previously. However, how the proteome profile changes due to this acute trauma has yet to be fully understood. To characterize the early proteome events induced by this trauma we compared proteome in muscle dialysate co… Show more

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Cited by 14 publications
(12 citation statements)
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References 44 publications
(54 reference statements)
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“…The proteins were reduced by incubating in 25 mM dithiothreitol for 15 min and alkylated 75 mM iodoacetamide for an additional 15 min. The samples were diluted 8× with 25 mM ammonium bicarbonate and filtered by 3 kDa Amicon spin-filter before digestion with trypsin (1:25, w / w trypsin/protein) [ 10 ]. The digested peptides were dried in a speed vacuum concentrator, reconstituted in 0.1% of formic acid in MilliQ water, and subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis.…”
Section: Methodsmentioning
confidence: 99%
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“…The proteins were reduced by incubating in 25 mM dithiothreitol for 15 min and alkylated 75 mM iodoacetamide for an additional 15 min. The samples were diluted 8× with 25 mM ammonium bicarbonate and filtered by 3 kDa Amicon spin-filter before digestion with trypsin (1:25, w / w trypsin/protein) [ 10 ]. The digested peptides were dried in a speed vacuum concentrator, reconstituted in 0.1% of formic acid in MilliQ water, and subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis.…”
Section: Methodsmentioning
confidence: 99%
“…Reversed-phase chromatography was performed using the EASY-nano LC II, as described previously [ 10 ]. The injection volume was 4 μL; mobile phases consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B).…”
Section: Methodsmentioning
confidence: 99%
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“…Database searches and label-free quantification. Database searches and label-free quantification (LFQ) were performed as described previously [80][81][82] . Raw files were searched using Sequest HT in Proteome Discoverer (Thermo; version 1.4.0.288) and X!…”
Section: Lc-ms/ms Analysismentioning
confidence: 99%
“…Commonly used techniques to investigate tissue proteomes include liquid based proteomic approaches, e. g. 2D gel electrophoresis or liquid chromatography (LC) based mass spectrometry 11 , 12 . However, these techniques do not enable a direct correlation between differentially expressed protein profiles and the tissue histology.…”
Section: Introductionmentioning
confidence: 99%