Structure/Folding

Differential Scanning Calorimetry (DSC) – Microcal VP-DSC

Used for the precise analysis of thermally induced structural transitions in proteins, DNA, RNA, micellar complexes, and other biomolecular systems.

Typical Applications:

• Stability and folding studies of protein and nucleic acids.
• Assessment of the effects of mutations on protein stability.
• Characterization of nucleic acids, lipids, membranes, and micellar systems.
• Measurements of the effect of ligand binding on protein stability. This allows the determination of the affinity of a very tight binding, with K_A up to approximately 10²⁰ M^-1

Differential Scanning Fluorometry (nanoDSF) – Tycho Nanometer

Used for the precise analysis of thermally induced structural transitions in proteins, DNA, RNA, micellar complexes, and other biomolecular systems.

Typical Applications:

• Batch-to-batch comparisons and assessment of the protein condition after storage.
• Assay development, screening buffer formulation conditions, etc.
• Protein functionality validation and detection of ligand binding.

Circular Dichroism (CD) – Applied Photophysics Q100 with Autosampler

CD spectroscopy measures the difference in the sample absorption of the left and right circularly polarized light. For the randomly oriented samples, such as isotropic solutions, this difference will be observed for the chiral solute molecules.

Typical Applications:

• Characterization of the secondary structure of proteins, peptides and nucleic acids.
• Detection of the interaction-induced conformational changes in proteins and nucleic acids.
• Determination of the effect of mutations on the protein structure.
• Study of the conformational stability of macromolecules at varying temperature, pH or denaturant concentrations:
  • Multiwavelength thermal unfolding measurements can be performed to simultaneously obtain the unfolding temperatures as well as the structural information on the intermediate and unfolded states from the 3D unfolding data.
  • Precise multi-point chemical unfolding studies can be performed automatically using the pipetting robot and Q100 autosampler.

Separation

Flow-Field Fractionation and Size Exclusion (FFF/SEC) Optilab

Molecular separations and measurements of molecular mass, size, and shape of macromolecules in solution.

Typical Applications:

• Measurements of the native molecular mass and size distributions of proteins and various macromolecules.
• Size separations in the 1 nm to 1000 nm range with no stationary phase.
• Oligomerization, interaction, and aggregation studies.
• Analysis of protein conjugates, micelle-solubilized membrane proteins, protein-nucleic acid complexes, nanoconjugates.
• Measurements of the second virial coefficient, conformation, and polymer branching.

Molecular Mass

Multi-Angle Light Scattering (MALS)- Optilab and Dawn

Molecular separations and measurements of molecular mass, size, and shape of macromolecules in solution.

Typical Applications:

• Measurements of the native molecular mass and size distributions of proteins and various macromolecules.
• Size separations in the 1 nm to 1000 nm range with no stationary phase.
• Oligomerization, interaction, and aggregation studies.
• Analysis of protein conjugates, micelle-solubilized membrane proteins, protein-nucleic acid complexes, nanoconjugates.
• Measurements of the second virial coefficient, conformation, and polymer branching.

Analytical Ultracentrifugation (AUC) – Beckman Proteomelab XLI

A gold-standard biomolecular analysis technique. A wide range of AUC-based methods are available for the analysis of interactions, oligomerization, composition, aggregation, membrane proteins, conformational changes, etc.

Typical Applications:

• SV-AUC:
• Oligomerization and interaction studies.
• Analysis of sample composition, aggregation, presence of trace components.
• Analysis of conformational changes.
• Analysis of viruses and nanoparticles.
• SE-AUC:
• Measurements of interaction and oligomerization affinities.
• Analysis of membrane proteins.

Mass Photometer (MP) – Refeyn OneMP

Native state, label-free and immobilization free single-molecule mass analysis in solution.

Typical Applications:

• Determination of sample heterogeneity, purity, polydispersity.
• Oligomerization analysis.
• Measurements of biomolecular interactions (K_D).
• Analysis of proteins and protein complexes, DNA, RNA, and lipids.

Binding Affinity

Microscale Thermophoresis (MST) – Nanotemper Monolith NT.115

Challenging biomolecular interactions measured in solution using microliter-volume samples in a capillary format. A wide range of ligands and affinities can be analyzed in various buffer conditions, including cell lysates and plasma.

Typical Applications:

• Measurements of a wide range of biomolecular interactions in solution.
• Binding measurements in challenging conditions, including cell extract, plasma, and other bio-liquids.

Fluorescence – PTI/Horiba QuantaMaster

A well established and universal method to study molecular interactions, conformation, stability, and to perform various biochemical assays.

Typical Applications:

• Studies of macromolecular interactions and ligand binding by detecting changes in the fluorescence intensity, anisotropy, or in the extend of the energy transfer (FRET).
• Structural studies of proteins and nucleic acids (tertiary structure changes of proteins, bending of DNA, etc.) by measuring changes in the intrinsic fluorescence, quenching or FRET.
• Studies of conformational stability of macromolecules at varying temperature, pH or denaturant concentrations.
• Measurements of metal ions concentrations, pH values, and other parameters in living cells, using fluorescent indicators.

Isothermal Titration Calorimetry (ITC) – iTC200

Label-free and immobilization-free studies of biomolecular interactions in solution.

Typical Applications:

• Native conditions binding affinity measurement without any labeling or immobilization.
• Applicable to a wide range of biological systems, including protein, DNA, RNA, and small molecule interactions.
• ITC is particularly well suited for the study of non-covalent interactions between biopolymers and small ligands, such as nucleotides, metal ions, etc.
• Enzyme kinetics measurements.
• Full thermodynamic characterization of binding.

Microfluidic Diffusion Sizing (MDS) – Fluidity One-M

Measurements of interaction affinity, stoichiometry, ligand concentration, and molecular size in native conditions, including serum, plasma, and cell lysate.

Typical Applications:

• Measurements of binding affinity in protein-protein and protein-nucleic acid interactions.
• Measurements of interaction stoichiometry.
• Measurements of the active target concentration in complex media (plasma, serum).
• Measurements of molecular size (free and bound probe).

Binding Kinetics

Bio-Layer Interferometry (BLI) – Octet RED96

High-throughput, kinetic binding analysis using the 96-well plate format and eight parallel detectors. Ready-to-use biosensors for antibodies, as well as for biotinylated, His- and GST tagged proteins.

Typical Applications:

• Measurements of binding kinetics and affinity.
• Off-rate ranking.
• Epitope binning.
• Antibody and protein concentration measurements.
• Automated, quantitative ELISA assays.