Biochemistry Research Facilities & Shared Equipment
The Department of Biochemistry has several pieces of shared equipment and shared research facilities that are available for use.
About the BIAcore S200 SPR & BIAcore 3000 Instruments
BIAcore S200 SPR
The Biacore S200 SPR instrument can measure interactions of various sample types, from low molecular weight drug candidates to high molecular weight proteins (also DNA, RNA, polysaccharides, lipids, cells, and viruses) in various sample environments (e.g., DMSO-containing buffers, plasma, and serum).
- Fragment screening and LMW drug discovery
- Kinetic and affinity determination
- Competition assays
- Epitope mapping of antibodies
The high sensitivity and low baseline noise of the Biacore S200 facilitate reliable affinity, kinetic, and fragment-screening data. The Biacore S200 is optimized for small molecule library screening, with automated software to evaluate non-specific binding, identification and prioritization of binders, and calculation of association (ka) and dissociation (kd) rate constants and affinity (KD). The Binding Level Screen function provides a rapid overview of small molecule libraries, automatically identifying fragments above a defined cut-off level. The predefined template for Binding Level Screen was developed explicitly for a 384-well plate format, allowing 384 compounds to be screened in less than 16 hours.
The S200 software also offers a range of tools for kinetic analysis, including analysis of single-cycle kinetics where several concentrations of an analyte are injected in the same cycle. Multicycle injections can still be analyzed, but by eliminating surface regeneration between injections, single-cycle kinetics simplifies the analysis of targets that are unstable or difficult to regenerate. Single-cycle kinetics also reduce analysis time. By utilizing the predefined templates for kinetics, thirty different analytes can be run in as little as 16 hours.
The Biacore S200 also simplifies competition studies to validate small molecules' interaction site with the capability to use ABA injections. The ABA injection mode allows two different solutions to be injected over the surface in the same cycle consuming much less inhibitor than traditional SPR experiments. In addition, the ABA injection feature can help identify ternary complex formation with multiple ligands.
The Biacore S200 instrument is housed in the Biochemistry Department. It is available to all Medical College of Wisconsin faculty and staff who are trained and can demonstrate proficiency on the instrument. Training and consultation are available on an appointment basis.
For more information on SPR, please see the following links provided by the manufacturer, Cytiva:
- Main Biacore page: Get started with SPR-interaction analysis
- Free on-line course: Get started with Biacore SPR assay
- Sign-up for Cytiva’s SPR seminar series: Biacore Online Spring Seminar Series 2021
The Biacore 3000 instrument remains available. However repairs will no longer be available as of September 2021. The BiaEval software for Biacore 3000 data will remain available for users.
The BIAcore 3000 instrument integrates surface plasmon resonance (SPR) technology with a microfluidics system to monitor molecular interactions in real time at concentrations ranging from pM to mM. This label-free technology can detect a wide range of molecular masses from 180Da to >1000kDa. The high sensitivity and high through-put capabilities allows for the detection of drug-protein, hormone-protein, protein-protein, DNA-protein, carbohydrate-protein, and lipid-protein interactions. The ability to interface with mass spectrometers provides discovery-based research in proteomic studies. For more information about SPR technology, theory, and applications, please see the BIAcore Service Presentation (PDF).
The BIAcore 3000 instrument is housed in the Department of Biochemistry and is available to all Medical College of Wisconsin faculty and staff who have been trained and demonstrate the ability to use microfluidic-based instrumentation. Training and consultation are available on an appointment basis.
Usage Fees: (Note: A training period of no less than 4 hours is required before you can work unassisted on the instrument. BIAcore chips and special reagents are not included in the fee.)
Academic Users: $50/hour | Industrial/Non-Academic Users: $75/hour
Academic Users: $12.50/hour | Industrial/Non-Academic Users: $40/hour
Consultation (experimental design, data evaluation)
Academic Users: $50/hour | Industrial/Non-Academic Users: $75/hour
For more information contact:
Nancy Dahms, PhD
(414) 955-4698 | firstname.lastname@example.org
Richard Bohnsack, MS
(414) 955-4699 | email@example.com
Biomolecular NMR at MCW
600 MHz NMR spectrometer. Cryoplatform is visible to the left of the magnet, RF console and workstation to the right.
The NMR Facility is an interdepartmental research service unit located in the Biochemistry Department. High-field NMR spectroscopy is a powerful technique for the study of biomolecular structure and dynamics. The facility provides service for routine 1D and 2D NMR methods, and can also provide consultation and collaborative assistance with the acquisition and analysis of multidimensional, multinuclear protein NMR spectra. The facility operates two Bruker 600 MHz and one 500 MHz NMR spectrometers, each equipped with 1H/13C/15N cryoprobes for enhanced sensitivity in biomolecular applications. In addition, a Bruker 300 MHz NMR spectrometer is available for routine analytical NMR of small molecules. For some long-term projects, the facility provides training for instrument operation and data analysis to investigators and research personnel. The facility operates on a fee-for-service basis and is open to faculty of the Medical College of Wisconsin and outside researchers.
The usage fee for internal users is $7.00 per hour for the 600 MHz NMR spectrometer, and $8.00 per hour for the 800 MHz Mass Spectrometer.
Macromolecular X-Ray Crystallography Facility
The department houses state-of-the-art instrumentation dedicated to Structural Biology research. The facility includes chromatographic systems for protein purification, an in-house X-ray diffraction core and an automated crystallization system for high-throughput screening and optimization. High-end computer workstations have been set up for 3-D graphic visualization and crystallographic analysis. The X-ray facility is located on the second floor of the Translational Biomedical Research Center (TBRC). The facility houses an X-ray diffraction system consisting of a Rigaku R-AXIS IV++ image plate detector system and MicroMax 007 generator equipped with Osmic confocal mirrors and an X-treme crystal cryocooler. The crystallization system includes a Hamilton STAR for solution making and a Phoenix equipped with a CrysCam for nanoliter crystallization and visualization. A fully automatic crystal incubator/imager (crystallization hotel) will soon be added to enhance throughput capacity. The facility is open to faculty members of the Medical College of Wisconsin. Various levels of training are available and collaborative arrangements can be made to scientists both inside and outside of the MCW community.
Biochemistry Shared Research Instrumentation
The Biochemistry Department maintains several instruments for isolation and physical characterization of biomolecules and detection of their interactions. All are located on the second floor of the TBRC and include:
Jasco J-710 Circular dichroism spectropolarimeter
The Jasco J-710 circular dichroism (CD) spectropolarimeter is equipped with a thermally regulated sample compartment. Monitoring of the far-UV and/or near-UV CD spectra can provide valuable information about the secondary structure, thermal stability, or conformational state of a protein.
Contact: Blake Hill, firstname.lastname@example.org
MicroCal VP - Isothermal Titration Calorimetry
The MicroCal VP-ITC is capable of measuring heat evolution as little as 0.4 nanoJ/sec. This instrument is suitable for the studies of protein-ligand and protein-protein interactions and provides the biochemists with reliable measurements of binding constants in the range of 103-109 M-1 as well as the enthalpy and stoichiometry of interactions. ITC is a preferred technique to demonstrate the interaction between newly discovered binding partners in vitro.
Contact: Brian Smith, email@example.com