Instrumentation
0077 Roy J. Carver Co-Laboratory
515-294-6473
12:00-1:00 Closed for lunch
By appointment/reservation.
Facility tours - Please contact Siquan Luo at 515-294-6473 or via
To analyze proteomes in the face of these challenges requires sophisticated technology. The new Proteomics Facility offers outstanding instrumentation to resolve complex proteomes and identify the component proteins. The Proteomics Facility will specialize in "differential proteomics," analyzing the differences in profiles of complex protein mixtures from different genotypes, environmental conditions, time points over a time course and so forth. Gel-based methods for resolving proteomes will utilize two dimensional difference gel electrophoresis (2D-DIGE) in which two samples can be run in the same gel for more accurate comparison. The thumbnails below will take you to bigger pictures.
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| Variable Mode Imager | ABI Q-Star XL Mass Spectrometer | High pressure liquid chromatography | Spot Handling Workstation | Surface Plasmon Resonance Instrument |
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To complement gel-based methods, multidimensional-chromatographic separations are available to expand the coverage of the proteome. These methods rely on different methods of protein separation than gel-based methods and, therefore, can often detect different proteins. Isotope-coding methods will be used for comparing and quantifying proteins in complex samples employing multi-dimensional chromatography approaches. Isotope coding, like 2D-DIGE methods for gels, allows two samples to be run in the same chromatographic separation, allowing for better quantification.
For both gel-based and chromatographic separations, proteins will be identified, when possible, by searching databases with both MS and MS/MS data using Mascot software. For organisms which do not have complete protein databases, de novo sequences from trypsin fragments can be obtained from MS/MS data. Digestion with other proteases will also be available to increase coverage when required.
In addition to differential expression surveys, analyses of the multiple forms of single protein species will be conducted. Many proteins are modified, for example, by cleavage, phosphorylation, or glycosylation, and these modifications often change the role of the protein in the cell or regulate its function. MS-based methods will be made available for the identification and mapping of post-translational modification of proteins. Differential expression analysis for identifying changes in post-translational modification patterns within the proteome will be available (phosphoproteome analysis, glycoproteome analysis etc.) Future development of the Proteomics Facility will address protein-protein interactions and the interaction of proteins with other molecules.