Rank buffer, salt, pH, additive, and ligand conditions by their effect on protein thermal stability using SYPRO Orange differential scanning fluorimetry, identifying conditions that maximize stability for crystallization and cryo-EM sample preparation.
Measure the absolute, conformation-independent molar mass and oligomeric state of a purified protein in solution by coupling size-exclusion chromatography to multi-angle light scattering and refractive-index detection, distinguishing monomer, defined oligomers, and aggregates.
Rapidly assess the homogeneity, oligomeric state, and structural integrity of a purified macromolecular complex by negative-stain EM, generating a low-resolution 2D characterization that gates whether the sample is suitable for cryo-EM.
Identify initial crystallization conditions for a freshly purified, monodisperse soluble protein by screening 96 sparse-matrix conditions in sitting-drop vapor diffusion, yielding diffraction-quality lead conditions for optimization.
Produce vitrified cryo-EM grids of a purified protein complex with thin, even ice and well-distributed particles in random orientations, suitable for high-resolution single-particle data collection.