The Gehrke Proteomics Center accepts any non-hazardous, non-pathogenic and non-radioactive samples for analyses. These samples can be whole organism (cell pellets, seedlings, etc.), tissue (biopsies, roots, leaves, etc.) or purified protein (cell lysate, precipitated protein, IPs, gel bands/spots).
We have standard operating procedures for each type of sample submission, but we welcome discussion of the goals of the experiment prior to initiation of a project (please contact Dr. Brian Mooney at mooneyb@missouri.edu).
Directed Analysis MS Lab
The Directed Analysis MS Lab is part of the Proteomics Center and is hosted by the Department of Biochemistry in Schweitzer Hall. We aim to assist campus researchers with quantitative analysis via liquid chromatography coupled to mass spectrometry (LC/MS). Our current instrumentation is used to quantify known compounds, for which standards are available. If you are interested in having experiments/analyses done in the lab, please contact our staff to discuss your project. More about the Directed Analysis MS Lab.
Global quantitative proteomics by mass spectrometry
LC-based separation of complex samples followed by simultaneous identification and quantitation of proteins is achieved using high-coverage accurate mass instruments (Bruker timsTOF-Pro and Pro2). We allow client-access to software for subsequent quantitative analyses, but our staff can also perform these analyses as fee-for-service. (Example data shown above.)
Targeted quantitative proteomics by LC-MRM
LC-based separation of complex samples followed by targeted quantitation of specific peptides/proteins using the Xevo triple quadrupole (QQQ). The instrument is capable of sensitive quantitation of proteins (e.g. biomarkers) or peptides (e.g. phosphorylation site occupancy).
Mass spectrometry-based identification of proteins
Identification of proteins is performed by obtaining data on trypsin-digested peptides and comparing these against public databases using MASCOT or Sequest software. Increasingly complex samples are analyzed by liquid chromatography mass spectrometry on the LTQ Orbitrap or Agilent 6520 QTOF.
Intact mass analyses
Intact mass (molecular weight) of purified proteins is obtained using our instrumentation and incorporates protein (C8 columns) LCMS for accurate mass intact protein analyses.
Post-translational modifications
Fluorescent stains and 2D gel electrophoresis allows specific imaging of phospho-proteins (ProQ-Diamond) and glyco-proteins (ProQ-Emerald). Additionally, post-translational modifications can be mapped by mass spectrometry. Mapping sites is not trivial, but we provide this service through extensive consultation with the client prior to, and on completion of, the analyses.
Two-dimensional electrophoresis
We perform high resolution large format (20x24cm) 2D-gel electrophoresis on any non-hazardous biological samples. The Center will accept whole organism, cell or tissue or purified protein samples. We are also capable of performing Western blots.
Image analysis
We can capture 16-bit TIFF images of 2D gels. Gels can be analyzed in-house (by staff or the client) using ImageMaster Platinum and/or DeCyder programs, the latter for analysis of DIGE (Cy dye-labeled) gels.
Center staff support
We emphasize our full-service aspect by involving staff and investigators at all times during sample processing. We facilitate experimental design, data analysis, manuscript figure/methods preparation, grant application figures, methods and facilities description preparation. Additionally, investigators can be trained by staff on sample prep, software and even instrumentation if, for example, a project requires extensive mass spectrometric analyses.