Mesecar Lab-The University of Illinois at Chicago (UIC)

Space, Instrumentation and Equipment that are Part of the Mesecar Lab:

General Wet Lab Space

We have three standard laboratory modules that are 565 sq. ft. each.  We have a fourth module for analytical instrumentation which is also 565 square feet.  All four modules, totaling approximately 2300 square feet, are adjoining and accessible via internal doorways. In addition, we have a separate room (250 square feet) that is used as a high throughput screening facility that houses our Tecan robot, Tecan plate reader, and Biorobotics colony picker.  We also have approximately 450 square feet of computational facilities that houses all of our SGI and Linux workstations and other computers.  In addition, we has space in shared cold rooms, warm rooms, and common rooms for freezers etc. In addition, we have access to autoclaves, dishwashers and other common equipment in the Center for Pharmaceutical Biotechnology.

Computational Resources and Computers

We have 18 PC's running Windows 2000/NT/XP, 10 SGI workstations running IRIX6.2, 1 dual processor Dell workstation under RedHat 7.2 Linux, 1 Linux workstation, 1 SGI 2100 Multiprocessor. We recently purchased a dual-processor Mac G5 X-Serve with 2.8 Terabytes of RAID Harddrive storage.  Our Center also has a shared Molecular Graphics Facility with commercial programs such as Felix, Sybyl (Tripos) with several modules, and InsightII, Discover, DelPhi, Xplor, Quanta, Catalyst (MSI), the Cambridge database, and associated programs.  Academic software includes Xeasy, Amber, Autodock, Grasp, Mopac, Macromodel, and Grid.  Crystallographic software includes HKL (Denzo, Scalepack), CrystalClear (MSC), CrystalStructure (MSC), MOSFLM, CNS, the CCP4 suite, SOLVE, O, Quanta, XtalView, Raster3D, Arp-Warp, Shake-n-Bake, SHARP, etc. We also have access to and use the BioE/RRC Computing Facility that consists of a 60 Node Linux Cluster (Pentium III, 1 GHz, each with 512Mb-1Gb memory) with 1.2 Terabyte storage.  We have installed many of the crystallography packages on this system.

Spectroscopy

Two Varian/Cary 50 UV/Vis spectrometers equipped with thermostated multi-cuvette(18) transports, PC-NT computers, and data analysis software are used for most of our visible and UV spectroscopy studies.  We also have two Gilson UV/Vis spectrometers with  thermostated 4-cell cuvette holder and chart recorders.  A research grade Varian Eclipse cuvette-based fluorescence spectrometer equipped with Peltier temperature control, automated polarizers, and an attachable 96/384 well microplate reader is used for our fluorescence and fluorescence polarization measurements. A Molecular Devices SPECTRAmax Plus 384 microplate reader with temperature control and PC-NT computer and software are used for routine UV/vis microtiter plate assays.  A JASCO Model 710 CD spectrometer connected with a Peltier temperature controlled sample compartment that are all interfaced to a PC with data analysis software are used for CD spectra and melting point determinations.

Protein Purification

We have fully automated Bio-Rad Biologic HR and Pharmacia AKTA FPLC systems, a Pharmacia AKTAprime, and a Bio-RAD LP low pressure chromatography systems for routine purification of proteins. 
We also have a Varian HPLC, Bio-RAD Rotofor preparitive isoelectic focusing system, Bio-RAD prepartive electrophoresis system, and numerous columns and resins.

Electrophoresis

Pharmacia PhastGel System, a Kodak Digital Electrophoresis Analysis and Documentation System and software, numerous PAGE and agarose analytical electrophoresis apparatus, BioRAD gel-dryers and pump.

Recombinant DNA and General Equipment

Two fully automated TECHNE thermocyclers, automated sonicator with temperature control, BioRAD electroporator, Sorvall preparative and micro centrifuges, New Brunswick Model 2000 10-Liter fermentor, and standard equipment such as pH meters, analytical balances, shakers, and incubators.

TECAN High-Throughput Liquid Handling Robot 

We recently puchased a Tecan Freedom Evo 200 liquid handling robot that is equipped with three robotics arms that are capable of simultaneous 8-syringe/position and 96/384-syringe/head manipulations via the TeMO 3/3 liquid-handling system.  We also have a Tecan Genios Pro multi-mode plate reader directly interfaced with the robot so that high-throughput screening and assays can be performed.  The plate reader has optics for both top and bottom reading and is capable of measuring UV/Vis, florescence intentsity and polarization, and luminescence.  The plate reader has full temperature control and is fitted with a single liquid injection port that allows for the initiation or quenching of reactions directly within the reader.  The robot is very versitile and can be used for compound library construction and dispensing, high-throughput screening assays, setting up sitting-drop crystal trays for protein crystallization, plasmid preparations, and other molecular biology manipulations etc.

BioRobotics BIOPICK^TM (Apogent Discoveries)-Flexible Colony Picking system. 

This system allows for the processing of up to 1,000 colonies per hour unattended with complete plate handling and automated plate filling.  The system can image, discriminate and pick from a 22 x 22 cm source plate with up to 10,000 colonies where the smallest colony can be 0.3 mm in diameter. 384 or 96 well plates, in addition to 96 deep well blocks can be used as destination plates.

Crystal Growth and Analysis

One Olympus SZ-60 and Two SZ-11 stereoscopes equipped with polarizers and a digital cameras, Hampton M6 automated incubator, numerous crystal growing solutions, chambers, manipulation tools, cryopreservation and supplies.

Additional access to other major equipment such as a DynaPro-801 dynamic light scattering instrument for measuring protein molecular weight and oligomeric or aggregation state, two EPR spectrometers, and preparative and ultra-centrifuges are available through shared facilities in our Center.  All of this equipment is located in the same building as our labs and offices.

Shared Facilities and Instrumentation at UIC & Argonne National Laboratory Commonly Used by Mesecar Lab Members

X-ray diffraction instrumentation is shared through the UIC Research Resources Center Macromolecular X-ray Diffraction facility.  The equipment is available to us on a regular basis and is paid for through an annual subscription fee of ~$5,000, depending on the actual annual cost.  This fee includes all service contracts, routine maintenance, parts, filaments etc. as well as all of our liquid nitrogen.  We currently have an Rigaku RU-200 x-ray generator, MSC mirrors system, Rigaku RAXIS-IIc imaging plate detector, Siemens HiStar multi-wire area-detector, and MSC and Oxford cryo-systems.  This facility is housed in our building and will be used extensively for structure elucidation. In 2003, we were awarded a grant from NIH/NCRR (1 S10 RR017744-01) to purchase an RAXIS 4++ x-ray detector and an X-Stream low-temperature cooling unit and to update the software and controlling hardware for the RaxisIIc.

We established the UIC/CPB/RRC Structure Determination Facility for determination of small molecule x-ray structures via a grant from NIH/NCRR (1 S10 RR-16848-01) to establish a campus x-ray facility.  This facility includes a 2.4KW x-ray generator, a Rigaku RAPID Area Detector, and a X-Stream low-temperature cooling unit.

The macromolecular x-ray instrumentation and the small molecule x-ray instrumentation are both housed in the Molecular Biology Research Building and are easily accessible by the Mesecar labs that reside in the same builiding.

  Synchrotron Access at the Advanced Photon Source. UIC is part of the South East Regional Collaborative Access team (SER-CAT) that has recently comissioned an insertion device beamline at the Advanced Photon Source (APS) at Argonne National Laboratory (ANL).  A second, bending magnet beamline will be available at SER-CAT within a year. This SER-CAT facility will guarantee the Mesecar Laboratory frequent and permanent access to synchrotron radiation, and will allow us to determine x-ray structures to the highest resolution possible as well as the ability to work with difficult samples. UIC is also an institutional member of the Consortium of Advanced Radiation Sources (CARS) at the Advanced Photon Source.  Within CARS is the BioCARS consortium that runs and maintains one insertion device and two bending magnet beamlines.  Our membership to CARS and hence BioCARS will also allow our lab frequent access to synchrotron x-rays.  We have extensively used all three of BioCARS beamlines for monochromatic, MAD, and Laue x-ray diffraction studies on a variety of projects. The APS SER-CAT and BioCARS facilities are located just 25 minutes away from UIC by automobile. Our membership in these consortiums will provide excellent opportunities for the continual training and education of students and post-docs in synchrotron radiation technology and x-ray data collection.

Mass Spectroscopy & Proteomics (Dr. Richard van Breemen Laboratory)

Within our Department of Medicinal Chemistry and Pharmacognocy, there are 6 mass spectrometers located in Dr. van Breemen’s laboratory.   The Mesecar group has an ongoing collaboration with Dr. van Breemen which gives us access to: Micromass Q-TOF Mass Spectrometer with Waters 2690 HPLC System; Micromass Quattro II triple quadrupole mass spectrometer, mass range of m/z 4,000, equipped with electrospray and APCI; a Hewlett-Packard HP5989B MSB mass spectrometer equipped with an electrospray, APCI, and HP1050 HPLC with autoinjector and photodiode array detector; ThermoFinnigan LCQ Deca Ion Trap Mass Spectrometer with  Surveyor HPLC System.  ThermoFinnigan TSQuantum Triple Quadrapole Mass Spectrometer with Surveyor HPLC System.  Applied Biosystems Voyager-DE Pro MALDI-TOF Mass Spectrometer.

NMR Spectroscopy (900 MHz, 800 MHz, 600 MHz)-UIC Center for Structural Biology

NMR Spectroscopy (400 MHz & 300 MHz)-Department of Medcinal Chemistry and Pharmacognosy

Isothermal Titration Calorimetery (Dr. Peter Gettins Lab)

Other Instrumentation Additional access to other major equipment such as a DynaPro-801 dynamic light scattering instrument for measuring protein molecular weight and oligomeric or aggregation state, two EPR spectrometers, and preparative and ultra-centrifuges are available through shared facilities in our Center.  All of this equipment is located in the same building as our labs and offices.