Analytical ultracentrifugation experiments play an integral role in the solution-phase characterization of biological macromolecules and their interactions. This unit discusses the design of sedimentation velocity and sedimentation equilibrium experiments performed with a Beckman Proteomelab XL-A or XL-I analytical ultracentrifuge and with a Beckman Optima AUC. Instrument settings and experimental design considerations are explained, and strategies for the analysis of experimental data with the UltraScan data analysis software package are presented. Special attention is paid to the strengths and weaknesses of the available detectors, and guidance is provided on how to extract maximum information from analytical ultracentrifugation experiments. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC.
Keywords: 2-dimensional spectrum analysis; AAV capsid quantification; Monte Carlo method; UltraScan; absorbance optics; analytical buoyant-density equilibrium; analytical ultracentrifugation; composition analysis; custom grid method; experimental design; intensity measurements; interaction studies; multi-wavelength analytical ultracentrifugation; parametrically constrained spectrum analysis; sedimentation velocity; solution studies; stoichiometry determination; van Holde-Weischet analysis.
© 2024 The Authors. Current Protocols published by Wiley Periodicals LLC.