Laboratories

 

The Case Center for Proteomics is comprised of 5000 square feet of laser, spectroscopy, spectrometry, cell culture and biochemistry laboratory space on the 9th floor of the BRB Building.

 

Biochemistry Facilities

 

HPLC and FPLC chromatography apparatus, microfuges, regulated water baths, gel dryer, vacuum pump, high voltage power supplies, sequencing gel units, protein and DNA electrophoresis apparatus, refrigerators, freezer, analytical balances, pH meter, micropipets, radiation safety equipment, glassware, pipettes, -70°C freezers and cold rooms.

 

Major Mass Spectrometry Equipment

 

The Proteomics Center has a full complement of mass spectrometry and proteomics facilities including:

• Thermo-Finnigan Fourier Transform LTQ mass spectrometer with 2 ppm resolution for top-down and bottom-up proteomics capabilities operated with Dionex capillary LC system; a 96-well autosampler and switcher is utilized with this system.
• GE/Amersham 2-D gel DIGE system with robotic spot picking.
• Applied Biosystems Q-star mass spectrometers. One has Agilent-nano-LC electrospray and one has ESI source.
• Thermo-Finnigan DECA XP-Plus and LTQ instruments, both with nanospray capability are available.
• Beckman Biomek FX robotic liquid handling systems is housed in the Proteomics laboratory with 96-well head and SPAN-8 flexible arm. This instrument is used internally by the center.
• prOTOF 2000 MALDI Mass Spectrometer, with attamole sensitivity and 5 ppm resolution.

 

Computation Facilities

 

This includes SGI workstations for viewing structures and larger scale server and computation systems for data analysis.

Administrative Support of the Case Center for Proteomics occupies a dedicated administrative suite adjacent to the center’s laboratory space on the 9th floor of the Biomedical Research Building at Case Western Reserve University’s School of Medicine.  The center’s support staff includes a Senior Administrator and Department Assistant to facilitate the purchasing of lab materials and supplies, maintain grant accounting and to assist in grant preparation.

 

Beamlines

 

The Center for Synchrotron Biosciences operates beamlines X9A, X28C, and X9B at the National Synchrotron Light Source for X-ray studies of various descriptions and U2B for infrared spectroscopy. The X-ray studies on X-9B and X-9A include x-ray absorption spectroscopy, macromolecular crystallography, and small-angle scattering. Each of the two beamlines is equipped with sagittal focusing monochromators and vertical focusing mirrors for high-flux bending magnet work (~5 x 1011 photons per second in a spot size of 0.25 x 0.3 mm from 5-15 keV). Both X9A and X9B beamlines have cryogenic cooling and CCD detection available for macromolecular crystallography (see below). Equilibrium and Time-resolved synchrotron footprinting is carried out on X28C, which has white beam.

 

Biochemical Laboratories

 

The Resource Center operates 2 biochemical laboratories and 2 cold rooms on the experimental floor for biological sample preparation and characterization and a spectroscopy lab for UV-VIS and infrared studies. Other equipment includes Beckman ultracentrifuge and Sorvall centrifuge; Cary 3 UV/Vis spectrophotometer; Anaerobic glove box, nanopure water system, ice maker, Cold room, pH mater, glassware, pipettors. These will be fully available for this project.

 

Special Equipment at NSLS for Footprinting

 

The X-28C beamline has a millisecond shutter for carrying out dose-response studies and Kin-Tec quench flow mixer for rapid kinetics studies. Special chambers for protein studies including flow cells for live cell exposure are available for this project.

 

Training

 

For additional information regarding instrumentation and services, please contact proteomicservices@case.edu.