UC Proteomics Laboratory (UCPL)

Ken Greis, PhD., Director


Standard Services

  • Comparative protein profiling:
    • Mini 2D gels
    • Comparative MS-based profiling using Label-Free and Isotope Tagging methods (SILAC, TMTs)
  • Protein identification by Mass Spectrometry
  • Characterization of Protein Complexes
  • Confirming and Mapping Protein Modification sites

Advanced Services/Collaborative Projects
  • Global Profiling of Protein Modifications (e.g. phospho, ubiquitin)
  • Complex MRM quantitation of targeted proteins
  • Enzyme assays and inhibitor screening by MS
 
Location & Hours:
The UCPL is located in the Vontz Center for Molecular Studies 1208-1216. Staff are typically available M-F, 9A-6P to accept samples, but please call 513 558-4057 to confirm.  First time customers should include a discussion with the director or associate director prior to preparing samples.

Services13
  • Mini 2D gel protein separation per gel

    Cost:

    130.00

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  • Mini 1D gel protein separation per gel

    Cost:

    95.00

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  • iTRAQ sample preparation and labeling per 4-plex sample

    Cost:

    410.00

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  • Preparative High pH RP separation of peptides with fraction collection per sample

    Cost:

    150.00

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  • Data Processing and Report Generation per hour

    Cost:

    45.00

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  • Research Staff hourly rate

    Cost:

    45.00

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  • MALDI-TOF/TOF and protein ID per sample

    Cost:

    60.00

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  • nanoLC-MS/MS for separation & fragmentation of peptides per sample

    Cost:

    175.00

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  • MALDI-TOF of intact protein per sample

    Cost:

    60.00

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  • Sample preparation by buffer exchange per sample

    Cost:

    30.00

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  • Protein Concentration by Pierce 660 assay per sample

    Cost:

    30.00

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  • Gel Stain (Silver Coomassie) per gel

    Cost:

    50.00

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  • Protease digestion and peptide recovery per sample

    Cost:

    30.00

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Equipment6
  • Two Sciex 5600+ Quadrapole-Tof Mass Spectrometer w/Eksigent nanoLC-Ultra 2D System

    One system is configured with an Eksigent nanoflow LC system and a nanospray interface primarily for proteomics application. It offers the hybrid capabilities for high resolution and high sensitivity profiling, characterization of compounds in complex mixtures, and also high selectivity and sensitivity for quantitation of compounds in complex mixtures by monitoring fragmentations transitions either in selected reaction monitoring (SRM) or multiple reaction monitoring (MRM) modes.  With attomole sensitivity, this system is optimized for quantitative proteomics profiling using label free (e.g. DIA, SWATH), isotope tagging methods (e.g. SILAC, iTRAQ) and quantitation of specific proteins (as peptide surrogates) from complex mixtures using SRM/MRM workflows.  The second system in coupled with the Dionex 3000 UHPLC units running at more conventional microflow rates for targeted quantitation analyses using SRM/MRM workflows for peptides and small molecule quantitation.

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  • Sciex 4800 MALDI-TOF/TOF MS


    The 4800 instrument is the primary instrument in the proteomics laboratory used for core services that require protein identification from single bands or spot from gels.  The attomole sensitivity and high repetition laser for fast data collection make it the ideal option for protein identification including protein complexes captured by affinity methods.  This instrument is also the primary system used for the rapid readout of enzyme assays and inhibitor screening studies.

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  • Beckman Coulter NX Liquid Handling Workstation

    Beckman Coulter NX Liquid Handling Workstation

    This system is use for samples preparation of enzyme reactions for MS-bases screening activities.

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  • Computational Systems and Software for Data Analysis include:

    1. 10 terrabyte general file server for automatic backup and archival of all proteomics data and reports.
    2. 8-processor MASCOT server for high throughput identification and quantification of proteins.
    3. 12-processor system running Sciex ProteinPilot software for relative quantitation of label-free or isotope tagged peptides
    4. 8-processor system running the Thermo Proteome Discoverer suite of data analysis software.

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  • Thermo Oribitrap Eclipse Tribrid mass spectrometer

    This newest system acquired via an NIH S10 instrumentation grant in October of 2019 is configured with a Dionex Ultimate 3000 RSLCnano LC and a FAIMS Pro High Field Asymmetric Waveform Ion Mobility Spectrometry Interface to allow for maximum separation and detection of peptides in complex mixtures.  This system is ideally suited for high throughput and high sensitivity comparative quantitative studies using Tandem Mass Tag (TMT) reagents or label-free quantitation methods.  With up to 5X better sensitivity compared to the Quadrupole-Tof systems in the lab, this system is used for global profiling of protein modifications (e.g. phosphorylation, ubiquitination) and for multiplex profiling studies of large number of samples.

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  • Thermo Oribitrap Fusion Lumos Tribrid mass spectrometer

    This high resolution mass spectrometer coupled with a Thermo Vanquish UHPLC system. This system is primarily dedicated to the development of new metabolomics profiling capabilities include steady state and metabolic flux analysis, as well as lipids, nucleosides, fatty acids and other metabolites.  With its high resolution accurate mass (HRAM), it is ideally suited for structural elucidate of small molecule via intact mass and fragmentation spectra for metabolomics needs.  With its rapid duty cycle and attomole sensitivity, it is dedicated for use to evaluate and quantify very complex sample mixtures for Metabolomics capabilities

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