Molecular Screening & Protein Expression Facility

The Molecular Screening and Protein Expression Shared Resource fosters collaboration by providing expertise in biochemical and cell-based assay development for high-throughput screening and compound profiling. Such assays enable researchers to identify small molecule compounds which interact with a target protein of interest. These compounds can then be used as tools to further study the target proteins function and signaling pathways in cells. The facility maintains a small molecule library of ~100,000 drug-like small molecules for high-throughput screening and provides access to liquid handling robotics and a multilabel plate reader. In addition, the facility also provides access to Biacore T200 SPR instrument for label-free binding kinetics and a high-content screening system. Guidance is provided for target justification, assay development, miniaturization, robotic automation, and adaptation to HTS-compatible, high-density microplate (384-well) formats. The facility provides production of recombinant proteins in insect cell systems and affinity purification of recombinant proteins upon request. The facility also maintains glycerol stocks of shRNA libraries, which can be accessed by investigators for use in target validation studies.

• Development and optimization of miniaturized biochemical, cell-based, high-content, and label free SPR assays
  
  
  • Receptor binding and functional assays, enzyme assays, protein-protein, protien-nucleic acid interaction assays
  • Cyototoxicity assays (caspase, mitochondria membrane potential, live cells), reporter gene assays, cell signaling assays
  • High-content cell-based assays in 96-well plates
  • Biophysical, label-free assays using Biacore, thermal shift, or ITC
    
    
• High-throughput screening and pharmacological profiling of small molecules
• Data analysis and interpretation
• Hit validation and quantitative assessment of compound activity required for post-screen advancement
• Consultation, training, project management, and grant preparation
• Recombinant protein expression in insect cells and other hosts on a case-by-case basis
• Affinity purification of recombinant proteins
• Distribution of shRNA library plasmids

• Janus Verispan 8-tip automated pipetting workstation
• Janus MDT automated pipetting workstation
• NanoHead MDT dispense tool (0.05-0.5 ul)
• EnVision multi-label microplate reader
• PerkinElmer Operetta High-Content Screening reader
• Biacore T200
• MicroFlo bulk reagent dispensers
• Amersham digital Western Blot system
• Libraries of compounds spanning various pathways and targets – contact the facility for more information
• BSL-1 and BSL-2 tissue culture suites