Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.2174/157016461104150121113824
Titel (primär) Magnetic bead-based proteome profiling using MALDI-TOF spectrometry in cardiac tissue from transgenic mice
Autor Baumann, S.; Gergs, U.; Schulz, N.; Thiery, J.; Neumann, J.
Quelle Current Proteomics
Erscheinungsjahr 2014
Department METABOX
Band/Volume 11
Heft 4
Seite von 229
Seite bis 236
Sprache englisch
Keywords Heart, MALDI-TOF MS, PP2A, protein profiling, transgenic mice
UFZ Querschnittsthemen RU3;
Abstract Development of a transgenic animal model always is followed by extensive characterization steps. Here, we intended to test and optimize a fast screening method to get general findings about the differential protein composition in the heart of wild type (WT) and transgenic (TG) mice. Therefore, we developed a protocol for magnetic bead-based separation (MB) combined with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDITOF MS) for protein profiling in plasma and cardiac tissue from TG and WT mice. We studied tissues from mice with cardiac specific overexpression of the catalytic subunit of PP2A, a model system for cardiac hypertrophy. EDTA-plasma or an extract of homogenized cardiac tissue (or skeletal muscle as control) were fractionated by hydrophobic interaction (MB-HIC C8) or weak cation exchange (MB-WCX) chromatography. MALDI-TOF MS spectra were generated in the mass range of m/z 1000-80000 using different matrices. The number of mass signals in the cardiac tissue extract was critically dependent on the use of the homogenization buffer, the residual blood contamination, and the surface modification of the magnetic beads. We noted different profiles in cardiac homogenates from WT compared to TG. As a control, the profile in skeletal muscle was not different between WT and TG. The results indicate that proteome profiling using MB-based sample preparation combined with MALDI-TOF MS is suitable for the proteome profiling in cardiac tissue of transgenic mice.
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=15849
Baumann, S., Gergs, U., Schulz, N., Thiery, J., Neumann, J. (2014):
Magnetic bead-based proteome profiling using MALDI-TOF spectrometry in cardiac tissue from transgenic mice
Curr. Proteomics 11 (4), 229 - 236 10.2174/157016461104150121113824