Techniques

ITC works by directly measuring the heat that is either released or absorbed during a binding event or an enzymatic reaction, and can simultaneously determine all binding parameters in a single experiment.

 

Technical data

Model – VP-ITC(MicroCal Inc)

Characteristics – Temperature range: 10-80 oC; Cell volume: 1.5 ml; Injection volume programmable (1-25 μL). Maximum volume of ligand added: 300 μl.

Applications – Thermodynamic characterization of intermolecular interactions in solution (ligand/biomolecule, non biological systems, nanoparticles, etc., where ligand includes small ligands, proteins, nucleic acids, carbohydrates, etc.) or enzymatic reactions. Quantification of binding affinity, drug selection and optimization, antibody titration, characterization of mechanism of action, determination of binding specificity and stoichiometry, measurement of enzyme kinetics, etc.

 

DSC is used to directly measure the intramolecular stability of structured macromolecules such as proteins and nucleic acids as well as the intermolecular stability of complexes such as oligomeric proteins, nucleic acid duplexes and lipid and detergent micellar systems, among other applications.

 

Technical data

Model – VP-DSC (MicroCal Inc)

Characteristics – Temperature range: -45 to 160 oC; Cell volume: 0.5 ml; Scanning rate variable (10 – 90 oC/h).

Applications – Thermodynamic characterization of protein and biopolymer conformational changes. Thermal stability of biomolecules (proteins, lipids, polysaccharides). Characterization of protein folding. Ligand identification. Optimization of pharmacological formations. Structure and stability of biopharmaceuticals. Determination of half-life of low molecular weight components.

 

Technical data

Model – Fluormax-4 (Horiba Jobin Yvon)

Characteristics – Two detectors (one is a calibrate photodiode reference detector to correct for intensity and temporal fluctuations in the source during excitation scans, the other is used in photon counting with signal linearity up to 2 x 106 CPS). Wavelength range: 220-600 nm for excitation and 290-850 nm for emission. Temperature range: 10 to 100 oC (peltier control system). Solid sample holder designed for samples such as thin films, powders, pellets, microscope slides and fibers.

Applications – Fluorescence spectroscopy (excitation, emission, synchronous scans). Temperature-wavelength scan (monitoring a range of wavelengths as a function of temperature). Kinetic scans.

Main application of these methods include: studying protein tertiary structure, protein unfolding as a function of temperature or added chemical denaturants, characterization of the effects of mutations on protein or nucleic acid structure and stability, studying macromolecule–ligand, macromolecule–macromolecule interactions, and kinetics processes. Micro-viscosity of membranes. Studying solid simples, films and fibers.

 

 

1. Manual glass-slide microarraying system

 

Technical data

Manufacturer: V&P Scientific, INC

Model: VP 478A / 472A / 470 / 475

Features: Enables the printing of duplicate microarray slides from 96 or 384 well plates with up to 768 spots per array. Samples can be spotted as duplicates, triplicates or more, with spot size ~500 microns in diameter (depending on slide surface), spot pitch 1,125 microns on horizontal axis and 750 microns on the vertical axis. The pitch can be changed easily for different spot sizes by skipping rows or columns.

 

2. Robotic Microarrayer

 

Technical data

Manufacturer: Arrayjet

Model: Sprint

Features: Enables the printing of up to 20 microarray slides from 96 or 384 well plates with up to 53,000 spots per slide. Samples can be spotted as duplicates, triplicates or more, with spot size ~100 microns in diameter (depending on slide surface). The robot is equipped with a temperature and humidity control system.

 

 

 

 

 

3. Microarray Scanner

 

Technical data

Manufacturer: Axon Instruments

Model: GenePix Autoloader 4200AL

Features: Four-color (blue, red, green and yellow) scanning enabling a broad choice of dyes, with automatic slide handling for up to 36 microarray slides. User selectable laser power (5-100% laser transmission in 1% increments) PMT gain and scan resolution (between 5 and 100 microns). Compatible with glass slides, membrane-coated glass slides and embedded arrays and a wide variety of samples (carbohydrate, protein, gene, tissue and cell arrays).

 

 

 

 

Technical data

Model – J-810 (Jasco)

Characteristics – Wavelength range: 163-900 nm. Temperature range: 4 -100 oC (peltier control system). Temperature scan rate (10 – 90 oC/h).

Applications – Measurement of ellipticity and absorbance at variable wavelength (spectra), time (kinetics) or temperature. Spectrum measurements (wavelength range: 163–900 nm). Kinetic processes (from second to minute timescale). Variable temperature (monitoring ellipticity at one-to-three different wavelength). Temperature-wavelength scan (monitoring a range of wavelengths, like in the spectrum measurement program, and a single wavelength as a function of temperature).

Main applications of these Circular Dichroism methods include: estimation of the secondary structure composition of proteins and peptides, characterization of protein–peptide unfolding as a function of temperature or added chemical denaturants, study of the effects of mutations on protein or nucleic acid structure and stability, methods for studying macromolecule–ligand, macromolecule–macromolecule interactions and chiral properties of organic compounds and kinetic processes.

 

Technical data

Model – OPTIMA XL-A (Beckman)

Characteristics – Maximum velocity: 45000 rpm. Temperature range (4 – 50 ºC). Detection system: UV-Visible (180–800 nm; up to three different wavelength in parallel). Number or rotor holes: 8 (7 for sample cells and 1 for counterbalance). Up to 21 samples per experiment in sedimentation equilibrium and 7 samples in sedimentation velocity.

Applications – Study of size, approximate overall shape, sedimentation coefficients and homogeneity of proteins and other biomolecules in solution. Detection and quantification ( species distribution, stoichiometry, reversibility, etc.) of interactions leading to formation of macromolecular complexes, including protein–protein, nucleic acid – protein, or receptor–ligand interactions.