Instruments – Keck Biophysics Facility

The Keck Biophysics Facility provides access to specialized instruments for sample characterization, absorbance, fluorescence, and circular dichroism spectroscopy.

	Analytical HPLC This Agilent 1100 series high pressure liquid chromatography system is equipped for high sensitivity sample analysis. Features: Diode-array absorbance detector from 190 to 950nm. Fluorescence detector. Peltier-thermostated auto-sampler (4-40 degrees C). Programmable automatic injector for 20- 900μl sample. Vacuum degasser. Peltier-thermostated column compartment (4-80 degrees C). Quaternary pump system using a high-speed proportioning valve. Note: The Keck Facility does not supply HPLC columns. Location: Cook Hall, 4106. NU Core ID: HPLC Agilent Technologies
	BLItz Bio-layer Interferometer The BLItz is a micro volume instrument for characterizing the kinetics of macromolecular interactions using bio-layer interferometry with low cost disposable sensor probes. Overview BLItz™ uses ForteBio’s Dip and Read™ label-free assays. These direct binding assays take place on a disposable biosensor made from a biocompatible matrix that is uniform, non-denaturing and minimizes non-specific binding. Only molecules that bind directly to the biosensor surface are detected, providing exceptional specificity for individual applications, even in crude media. The BLI (bio-layer interferometry) technology used by BLItz provides real-time data on protein interactions. BLItz emits white light down the biosensor, and then collects any light reflected back. Reflected wavelengths are affected by the thickness of the coating on the optical layer. Some wavelengths show constructive interference (blue), others show destructive interference (red). This interference is captured by a spectrometer as a unique spectral signature and is reported in relative intensity units (nm). Any change in the number of molecules bound to the biosensor causes a shift in the interference pattern that is measured in real time. This wavelength shift is a direct measure of the change in optical thickness (nm) of the biological layer. Biosensor Types For a list of all available biosensor types, please see here. Note that our facility has select sensor types (please check with us to confirm the specifics) available for purchase to users. An instrument is located on each campus. Location: Cook Hall, Rm 4106, Evanston Campus Location: Tarry Building, Rm 7-710, Chicago Campus - Not Available NU Core ID: Bio-Layer Interferometer (BLITZ) Evanston
	Differential Scanning Calorimeter The MicroCal (GE Healthcare) differential scanning calorimeter directly measures the enthalpy of a solution as a function of temperature. Applications of the DSC technique: Protein stability and folding. Protein engineering. Characterization of membranes, lipids, nucleic acids and micellar systems. Assessment of the effects of structural change and ligand binding on a molecule’s stability. Instrument Characteristics: Active cell volume ~ 0.5 ml. Non-reactive Tantalum 61™ cells for excellent chemical resistance. Fixed-in-place cells for reproducible ultrasensitive performance with low maintenance. Operating temperature range of –10ºC to +130ºC. Peltier element for precise temperature control. User selectable temperature scan rates (0ºC to 90ºC per hour upscans) and range for application versatility. Location: Cook Hall, 4106 NU Core ID: DS Calorim MicroCal \| VP-DSC
	Isothermal Titration Calorimeter The MicroCal (GE Healthcare) ITC200 isothermal titration calorimeter is a powerful tool for label-free analysis of macromolecular interactions in solution. By measuring the amount of heat released or taken up during each reaction in a titration experiment, the equilibrium binding affinity and the stoichiometry are derived; in addition, ITC is the only technique able to directly measure the thermodynamic parameters of binding (enthalpy, entropy and heat capacity changes) – providing therefore invaluable insights into the thermodynamic forces governing biomolecular interactions. The technique is suitable for analyzing protein-protein, protein-DNA/RNA interactions as well as interactions between biopolymers or synthetic macromolecules and small ligands. -Affinity constants from 100 to ~109 M-1 can be measured by ITC. -The sample cell volume is approximately 200 μl; minimum sample volume is 290 μl. -The syringe volume is 40μl. -The operating temperature range is 2°C to 80°C. Location: Cook Hall, 4118 NU Core ID: ITC200 MicroCal \| ITC200
	Affinity ITC The new Affinity ITC ( TA Instruments) is a powerful tool for measuring a wide variety of molecular interactions. It brings advanced engineering to all critical aspects of the measurement ensuring the highest quality ITC data. Features -AccuShot™ delivers the titrant to the right location for the best mixing -FlexSpin™ provides innovative slow speed stirring, efficient mixing and highest sensitivity -Fully automated, user-selectable system cleaning routines eliminate run-to-run contamination -Intelligent Hardware Positioning for precise reliable injections -Solid-state active heating and cooling for true isothermal temperature control -Low volume cells (190 µL) -Powerful ITCRun and NanoAnalyze for the most comprehensive suite of tools for method optimization, model fitting, batch analysis, graphing and data export. NU Core ID : Affinity ITC Contact Keck Biophysics for more information TA Affinity ITC
	NanoDLS pUNK The pUNK, or NanoDrop DLS, from Unchained Labs offers a fast and easy method to measure protein size, aggregation, and polydispersity. The smallest, fastest and easiest to use protein sizing system. Only 5 µL of sample required to determine: Percent aggregation. Polydispersity. Location: Cook Hall, 4118 NU Core ID: Punk More Information
	SEC-MALS-QELS The SEC-MALS-QELS system is composed of a size exclusion chromatography system (SEC) followed by a Wyatt DAWN HELEOS II multi-angle static light scattering detector, a Wyatt QELS dynamic light scattering detector, and a Wyatt T-rEx differential refractive index detector. System components: Wyatt DAWN HELEOS II – multi-angle (static) light scattering (MALS) detector. The molar mass and root mean square radius of the solute are determined by measuring the intensity of the scattered light – provided that the refractive index increment (dn/dc) of the solute/solvent system is known. Wyatt Optilab T-rEx – refractometer for measuring the absolute refractive index and its increment Wyatt QELS – quasi-elastic (dynamic) light scattering (QELS) detector. The diffusion coefficient/hydrodynamic radius can be determined from the temporal fluctuations in the intensity of scattered light HPLC – with size-exclusion columns (SEC), DAD absorbance, and fluorescence detectors In this set-up the sample components are separated according to size and characterized individually by static and dynamic light scattering, absorbance and fluorescence. Typical applications: For proteins and nucleic acids: measurements of molecular weights and radii, oligomerization, heterogeneity and solubility studies, characterization of macromolecular complexes in terms of size, overall shape and composition/stoichiomentry. For synthetic polymers: measurements of molar mass & mass moments, polydispersity, conformation/branching. For nanoparticles: solubility and size distribution measurements and composition /stoichiometry analysis for functionalized nanoparticles. More Applications Location: Cook Hall, 4106 NU Core ID: MALS QELS
	Octet K2 Bio-Layer Interferometer The Octet K2 System characterizes protein-protein and protein-small molecule binding interactions for early research and discovery. The multipurpose system also provides quantitative information by measuring active protein concentrations — even in complex mixtures like cell culture supernatants and lysates. Features: Sensitivity for Small Molecule Analysis The Octet K2 System enables the measurement of interactions between small molecules down to 150 Da in size, and their larger binding partners to determine accurate kinetics rates. Flexible Second Channel for Use as a Reference or More Samples Unique 2-channel design have dedicated spectrometers for simultaneous referencing or added capacity. Simple Setup for Protein and Antibody Characterization Template driven interface allows new users to get started quickly for protein and antibody characterization. Location: Cook Hall, 4118 NU Core ID: BLItz Bio-Layer Interferometer Octet
	Surface Plasmon Resonance The Reichert 4 SPR system is a four channel surface plasmon resonance spectrometer for label-free measurement of macromolecular association and dissociation kinetics. SPR is suitable for a wide range of interactants including proteins, nucleic acids, lipids, carbohydrates, small drugs and synthetic polymers and nanoparticles. The Reichert4SPR 4-Channel Surface Plasmon Resonance system allows users to accelerate and enhance the drug discovery process in the following ways: -Easy set-up and streamlined workflow. -Utmost sensitivity and maximum uptime. -Robust fluid handling system to run demanding assays. Features: 4 channel channel acquisition. Autosampler holding up to 96 HPLC vials or two 96 or 384 well plates. Controllable injection volume to conserve precious analytes. Optional sampling cooling for extended assays. Interaction temperature controllable between 10°C and 70°C. Detection of small molecule analytes down to ~100 daltons. Accessible range of binding affinities from 1 mM to 1 pM. Immobilization Techniques: Amine coupling to carboxylated planar monolayer or branched dextran sensor chips. Thiol coupling using a native or introduced cysteine. NeutrAvidin, streptavidin, or avidin capture of biotinylated molecules. Nickel NTA capture of histidine-tagged molecules. Protein A capture of antibodies. Lipid monolayer formation on a hydrophobic surface. Reichert provides an assortment of sensor chips to accommodate different immobilizations. Please see here for a complete list of available sensor chips. Location: Cook Hall, 4106 NU Core ID: SPR More Information

	Circular Dichroism Spectrophotometer Our state of the art Jasco J-815 CD instrument allows the simultaneous acquisition of fluorescence and CD data. Such experiments are particularly useful in assessing the extent to which protein unfolding and folding transitions are two-state in character. Select Applications: Protein conformational studies. Quantitative analysis of pharmaceuticals. Formulation studies. Protein folding studies. DNA/RNA interactions. Enzyme kinetics. Purity testing of optically active substances. Natural products chemistry. Accessories: Two photomultiplier tubes (165-900 nm and 600-1100 nm). Peltier sample holder (used with 1mm, 2mm, 5mm, or 10mm path length cuvettes) for thermal denaturation studies (4- 110 degrees Celsius). For more information about the instrument, please see here. Location: Cook Hall, 4118 NU Core ID: Circular Dichroism Spec. J-815
	Fluorescence Plate Reader The Molecular Devices Gemini EM Fluorescence/Chemiluminescence Plate Reader complements the facility’s fluorimeters PC1 and K2 by accommodating high throughput fluorescence experiments. Features: Dual monochromators, allowing the determination of optimal excitation and emission settings in 1 nm increments. Top and bottom reading options, allowing measurements for both solution and cell-based assays. Location: Cook Hall, 4118 NU Core ID: FL Plate Reader Gemini
	NanoDLS pUNK The pUNK, or NanoDrop DLS, from Unchained Labs offers a fast and easy method to measure protein size, aggregation, and polydispersity. The smallest, fastest and easiest to use protein sizing system. Only 5 µL of sample required to determine: Percent aggregation. Polydispersity. Location: Cook Hall, 4118 NU Core ID: Punk More Information
	NanoDrop 3300 Fluorimeter The Thermo Scientific NanoDrop 3300 fluorimeter acquires full range visible emission spectra from micro-volume samples in seconds. Features: Measurements require only 1-3 µL of sample. UV, Blue, and White LED excitation sources. Spectrometer recorded emissions between 400-750 nm. Fluorescent Profiler tool quickly determines appropriate excitation source, emission spectrum, and peak emission wavelength. Pre-configured methods for Nucleic acid quantification, Protein quantification, and for common fluorophores. Can measure multiple fluorophores in a single sample. Exports quantitative spectral data. Location: Cook Hall, 4106 NU Core ID: NanoDrop FL (Re-Opening)
	NanoDrop Spectrophotometer The Thermo Scientific NanoDroop 2000c acquires full range (190mm - 840mm) UV/Vis absorbance spectra from micro-volume or cuvette samples in seconds. Drop measurements require only 1-3 µL of sample or measure from standard 1 cm Cuvettes. Individual scans take less than 5 seconds. Exports full spectrum readings to excel. Pre-configured methods for: DNA. RNA. Peptides. Protein A280. Calculating sample purity ratios 260/280 nm and 260/230 nm. Cell culture readings 600nm. Location: Cook Hall, 4106 NU Core ID: NanoDrop Abs
	Photon-Counting Fluorescence Spectrometer The ISS PC1 spectrofluorimeter is a workstation for a wide variety of highly sensitive fluorescence studies. The PC1 has excitation/emission monochromators and optional polarizers suitable for fluorescence polarization/anisotropy and ratio measurements using both T- and L- configuration. Measurements that can be performed with the PC1: Corrected excitation and emission spectra and (new!) 3-D excitation-emission matrices. Corrected excitation and emission polarization (anisotropy) spectra and measurements at a fixed wavelength. Synchronous luminescence spectra. Slow kinetics (intensity, ratio, anisotropy, etc.), in time spans ranging from tenths of seconds to days. Kinetic capability in the millisecond range. Automated temperature scans. Automated titration experiments. The Keck Facility provides a collection of cut-on filters. Location: Cook Hall, 4118 NU Core ID: PC1 ISS \| PC1™
	UV/Vis DAD Spectrophotometer The HP (Agilent) 8452 diode array spectrophotometer is a basic analytical instrument for measuring UV/Vis spectra of solution samples in seconds. Detection range (190-820nm) is linear between 0.2-1.1 AU with a wavelength accuracy of 1-2 nm. Cuvettes: Quartz cuvettes are required for measurements from 190nm to 350nm. Glass cells and/or disposable plastic cells can be used from 350nm to 820nm. Note: If you need to measure a solid sample or measure in the NIR region, please consider the UV/Vis/NIR, Lambda 1050 instrument. Location: Cook Hall, 4106 NU Core ID: UV/Vis Spec Agilent Technologies HP8452
	UV/Vis/NIR Spectrophotometer The Perkin Elmer LAMBDA 1050 is a high performance UV/Vis/NIR double beam, double monochromator, ratio-recording spectrophotometer. A 150 mm integrating sphere accessory allows diffuse reflectance and transmission measurements. Instrument Features: Wavelength range 185 nm – 3300 nm. Photometric Range: 0.0002- 9 AU. Liquid and solid samples measuring capabilities. Sample thickness compensated detector optics. 3 detector module: photomultiplier R6872, 3-stage wide band Peltier cooled InGaAs and single-stage Peltier cooled PbS. Spectral bandwidth : 0.05 nm – 5 nm UV/Vis and 0.2 nm – 20 nm NIR. Programmable attenuators for high absorbance measurements. UV WinLab V6 operating software. Diffuse Reflectance and Diffuse Transmission measurements can be performed using the 150mm Integrating Sphere accessory. Select Applications of Lambda: Absorption Spectroscopy as a Powerful Technique for the Characterization of Single-Walled Carbon Nanotubes. APP_SpectroscopicInVitroSunscreenSPFValues. Reflectance Measurements of Materials Used in the Solar Industry. Measuring Absorptance (k) and Refractive Index (n) of Thin Films with the PerkinElmer Lambda 950/1050 High Performance UV-Vis/NIR Spectrometers. Simple Method of Measuring the Band Gap Energy Value of TiO2 in the Powder Form using a UV/Vis/NIR Spectrophotometer. Visible Reflectance Spectroscopy of Human Skin: the use of CIE Lab* Color Analysis for In Vivo Ethnic Skin Characterization. Characterization of Custom Light Sources using the LAMBDA 650/750/850/950/1050 UV/Vis and UV/Vis/NIR Research Spectrometers. Location: Cook Hall, Room 4106 NU Core ID: Lambda Spec (UV/VIS/NIR) PerkinElmer \| LAMBDA 1050

	Azure 600 Imaging System The Azure 600 gel imaging system allows visualization, imaging and quantitative measurements of macromolecules in stained gels. It’s large size also makes it ideal for imaging test tubes, petri dishes, plates, etc. Capabilities - Chemiluminescent imaging - Fluorescent Imaging -----Blue, Green, Red, and NIR epi-light sources (light from above) -----Trans-UV light source (light from below) -White Light imaging -----True color imaging -Trans- White light Features -9.1 MP camera for publication-ready images - Multiple detection modalities in a single instrument – Can mix and match filters with different light sources -Can auto-expose specific regions of image -High functional dynamic range for simultaneous detection of low & high abundance targets -No manual focusing regardless of gel size -Use with a wide range of fluorescent probes -Multiple targets can be probed on a single blot with up to three visible fluorescent channels -Bands can be excised without leaving the imager -DNA imaged with minimal damage -Fluorescent blot reimaging over weeks -High CCD sensitivity & speed improves target detection even when signals are faint or fading Analysis Software: AzureSpot Location: Cook Hall, 4106 NU Core ID: Azure 600
	Lyophilizer The Labconco FreeZone 4.5 Liter Cascade Benchtop Freeze Dry System is designed for light sample loads with low eutectic points including ones containing acetonitrile. Features: Removes a maximum of over 4 liters of water in 24 hours.* Includes brushed stainless steel and powder-coated steel exterior. Stainless steel collector chamber and coil. LCD that displays system operating parameters. Set-up parameters and alarm messages (choice of temperature in °C and ° F, choice of vacuum in mBar, Pa and Torr). 1/3 hp CFC-free refrigeration system. Red alarm light. LED vacuum and temperature wave graphs. Patented moisture sensor. Vacuum control/break valve. Rear-mounted RS-232 port. Automatic start-up switch for collector cool-down and vacuum pull-down with manual override switches. Retractable collector drain line. Power switch; rear-mounted electrical receptacle for vacuum pump connection (pump not included). 3/4″ OD vacuum connection; 3/4″ ID vacuum hose; and two clamps. 12-port stainless steel drying chamber. * Freeze drying rate will be lower for samples other than shell-frozen plain water. For optimum performance, room temperature should be 21° C (70° F) or colder. For more information, click here. Location: Cook Hall, 4118 NuCore ID: Labcono Lyophilizer
	Preparative Ultracentrifuge Sorvall MTX 150 Ultracentrifuge with an assortment of rotors. Features: Sample separation up to 1,048,000 × g. Versatile rotor capacities. Fixed angle or swinging buckets rotors. Vacuum conditions. Temperature control. Location: Cook Hall, 4106 NuCore ID: Preparative Ultracentrifuge More Information
	Real Time PCR The Bio-Rad iCycler collects and reports PCR amplification and denaturation data in real-time. Key features of the CFX96 Touch system include: -6 filtered LED’s and 6 filtered photodiodes in the 450-730 nm range. Can monitor SYBR Green or multiplex. -Monitoring of 96 wells with up to 5 fluorophores per well simultaneously. -Dynamic range of ten orders of magnitude for detecting specific sequences with excellent sensitivity and precision. -Updated software for easy set up and data analysis. -Results emailed directly to you. -Master Mix Calculator outputs a simple list of instructions and volumes for mixing reagents. -Easily creates normalized gene expression graphs. -Sample volumes 10-50 µl. Location: Cook Hall, 4106 NU Core ID: qPCR Bio-Rad iCycler Michael Zuker’s Algorithms Handbook
	Sapphire Multi-Mode Imager The Sapphire™ Biomolecular Imager is a next generation laser scanner which features NIR and RGB fluorescent, true chemiluminescent and phosphor imaging. Four solid state lasers (488, 520, 658 and 784 nm). Photomultiplier tube (PMT) for fluorescence and phosphor imaging, avalanche photodiodes (APD) for near-infrared imaging and a CCD sensor for chemiluminescent and visible imaging. Ultra-wide dynamic range for imaging and quantifying low and high abundance samples simultaneously. Image resolution down to 10 microns for high-quality image analysis. Advanced capabilities for documentation and quantitative analysis of gels, blots, plates, arrays, slides, tissue samples, small animal models, plants, and more. Analysis Software: Azure Spot Location: Cook Hall, 4106 NU Core ID: Sapphire Imager (Re-Opening) Please acknowledge the use of the Keck Biophysics Facility's Azure Sapphire Imager -funded by the 1S10OD026963-01 NIH grant when you include images or data acquired on the Sapphire in your publications. This explicit acknowledgment is required by the NIH.