Instruments

Instrumentation and services are available to all researchers inside and outside of the university.

Researchers from the Northwestern University and the University of Chicago are provided internal rates as all UIC users. All other users can access services but at an external academic or external non-academic rate, which is determined using subsidy and market rates.

 

All users must register their funds by generating an RRCAN before requesting service or scheduling equipment usage.

 

All users are responsible for their own laboratory safety training to be current during each use of the core facilities.

Biacore T200 (Surface Plasmon Resonance)

Biacore T200

Location:  Molecular Biology Research Building (MBRB) Room 3327

Description:  The Biacore T200 is a very advanced SPR instrument that can support real-time detection and monitoring of the biomolecule interactions such as proteins-proteins, proteins-peptides, proteins-DNA, and proteins-compounds. Our Biacore T200 can detect from large protein complexes (> 100 kDa) to a very small fragment compounds (< 100 Da), and it can provide quantitative information on binding specificity, binding affinity, kinetics, and concentration. This technique can also be applied to every stage of drug-discovery, which includes mid-throughput automated compound screening, hit confirmation and validation, hit characterization via kinetics, and mechanism of action.

Jasco 815 Circular Dichroism (CD) Spectrometer

Jasco 815 Circular Dichroism (CD) Spectrometer

Location:  CSB (Center for Structural Biology) Room 113

Description:  Circular dichroism measures the interaction of circularly polarized light with molecules.  Circularly polarized light interacts equally with non-chiral molecules so non-chiral molecules are not measured.  As the circularly polarized light passes through an optically active substance, its two circularly polarized components travel at different speeds and are absorbed in differing degrees.  The Jasco 815 CD spectrometer can measure conformation in chiral molecules.  CD spectra between 260 and approximately 190 nm can be analyzed for secondary structural elements: alpha helix, parallel and anti-parallel beta sheet, turn, and other.  The absence of regular structure results in zero CD intensity, while an ordered structure reveals a spectrum that can contain both positive and negative signals.

  • Characterization of protein secondary and tertiary structure
  • Studying the conformational stability of a protein at varying temperature, pH or denaturant concentrations
  • Determination of the effects of protein-protein interactions on conformation
  • Comparing the structures of protein vs. mutants or proteins expressed in different systems

Dynamic Light Scattering (DLS)

Dynamic Light Scattering (DLS)

Location:  CSB (Center for Structural Biology) Room 113

Description:  When monochromatic coherent light (laser light) interacts with particles in solution one observes a time-dependent fluctuation in scattering intensity.  This correlation arises from the movement of the particles in solution due to Brownian motion.  As the distance between the particles in solution fluctuates, the scattered light undergoes constructive and destructive interference in a time-dependent fashion.  This contains information about the time scale of the movement of the particles and thus can be correlated to size.

  • To estimate protein molecular weight
  • To observe protein oligomerization
  • To determine the quality of a protein sample for structural studies.  For example if the protein has aggregated.

Analytical Ultracentrifugation (AUC)

Analytical Ultracentrifugation (AUC)

Location:  CSB (Center for Structural Biology) Room 113

Description:  The Beckman ProteomeLab XL-1 measures concentration distributions of a sample in solution in a centrifugal field.  The application of centrifugal force causes a redistribution of the macromolecules and the formation of a concentration boundary that moves from the meniscus to the bottom of the cell over time.  A significant strength of technique is that the properties of native proteins are studied in solution -requiring no label, chemical modification or surface interaction.

The XL-1 is equipped with two optical detection systems.  The first is capable of reading the absorbance of a sample using a dual-beam UV/VIS spectrophotometer with monochromator.  The second detection system uses a laser interferometer that records the refractive index gradients.  The choice of the optical detection system depends on the sensitivity needed for the experiment, the concentration range to be used, the extinction properties of the system and the buffer properties.

Two methods used for running the analytical ultracentrifuge are sedimentation velocity and sedimentation equilibrium.  In sedimentation velocity, high rotor speed sediments macromolecules to the bottom of the cell.  The rate of sedimentation is dependent on the size and the shape of the protein.  Sedimentation velocity experiments generally run for 6 – 12 hours.   Sedimentation equilibrium experiments are run at lower rotor speeds where the process of sedimentation is balanced by diffusion.  When no change in the concentration distribution is detectable, sedimentation equilibrium is achieved.  The time required to establish sedimentation equilibrium is generally 1 -2 days with the entire experiment typically taking between 3 and 5 days.

  • Determine the molar mass of proteins and complexes
  • Assess the purity of a sample
  • Determine the number of species in a sample
  • Determine the stoichiometry of complexes
  • Analysis of self- and hetero-association
  • Determine binding constants (10-3– 10-8 M)
  • Characterizes synthetic polymers, biological macromolecules, viral particles, carbohydrates and nanoparticles

Isothermal Titration Calorimetry (ITC)

Isothermal Titration Calorimetry (ITC)

Location:  CSB (Center for Structural Biology) Room 113

Description:  The MicroCal VP-ITC is useful for the characterization of the thermodynamics of a binding reaction in solution.  In an ITC experiment, aliquots of a titrant (typically a protein, peptide or small molecule) are injected into the cell containing a macromolecule solution.  With each titration injection, the molecules interact and heat is either generated or absorbed.  The VP-ITC measures this heat of binding to determine the binding constants (K), reaction stoichiometry (n), enthalpy (ΔH) and entropy (ΔS). In addition, varying the temperature of the experiment allows the determination of the heat capacity (ΔCp) for the reaction.  Since the heat of binding is a naturally occurring event, the ITC does not require immobilization and/or modification of the reactants.  There are no limits on the protein or ligand size nor is the system dependent on the optical properties of the samples.  The major limitation of the ITC is that it requires relatively high concentrations of samples.

  • To monitor binding interactions, such as but not limited to: antigen-antibody, DNA-drug, receptor-target, protein-ligand or protein-protein
  • Determination of reaction stoichiometry
  • Measurement of binding constants
  • To measure the thermodynamic properties of binding – enthalpy, entropy and Gibbs free energy

Emulsiflex C5 (Cell lysis)

Location:  CSB (Center for Structural Biology) Room 113

Description:  The Emulsiflex C5 is a gas driven homogenizer designed for high-volume cell lysis.  Cells are resuspended in buffer and passed through the system under high pressure.  Unlike a French Press and Sonication, the volume of resuspended cells is virtually limitless, and heating of the sample is minimal.

  • Efficient lysis of bacterial cells
  • Only Biosafety Level One cultures are permitted

PerkinElmer Victor3V plate reader

Location:  Molecular Biology Research Building (MBRB) Room 3368

Description:  A general-use spectrophotometer with multiple detection capabilities:

  • Fluorescence intensity (top & bottom reading)
  • Fluorescence Resonance Energy Transfer(FRET)
  • Absorbance (UV to NIR range)
  • luminescence

Scheduling

Please email Hyun Lee (danielhl@uic.edu) to schedule a meeting or a training.  Once you become an approved self-user, you can do on-line scheduling by yourself using UICore.