Measurement of the temperature of the resting rotor in analytical ultracentrifugation

Rodolfo Ghirlando; Huaying Zhao; Andrea Balbo; Grzegorz Piszczek; Ute Curth; Chad A Brautigam; Peter Schuck

doi:10.1016/j.ab.2014.04.029

Measurement of the temperature of the resting rotor in analytical ultracentrifugation

Anal Biochem. 2014 Aug 1:458:37-9. doi: 10.1016/j.ab.2014.04.029. Epub 2014 May 4.

Authors

Rodolfo Ghirlando¹, Huaying Zhao², Andrea Balbo³, Grzegorz Piszczek⁴, Ute Curth⁵, Chad A Brautigam⁶, Peter Schuck⁷

Affiliations

¹ Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: rodolfo.ghirlando@nih.gov.
² Dynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892, USA.
³ Bioengineering and Physical Science Shared Resource, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892, USA.
⁴ Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
⁵ Institute for Biophysical Chemistry, Hannover Medical School, 30625 Hannover, Germany.
⁶ Department of Biophysics, The University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁷ Dynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: schuckp@mail.nih.gov.

Abstract

Accurate measurements of rotor temperature are critical for the interpretation of hydrodynamic parameters in analytical ultracentrifugation. We have recently developed methods for a more accurate determination of the temperature of a spinning rotor using iButton temperature loggers. Here we report that the temperature measured with the iButton on the counterbalance of a resting rotor, following thermal equilibration under high vacuum, closely corresponded to the temperature of the spinning rotor with a precision better than 0.2°C. This strategy offers an inexpensive and straightforward approach to monitor the accuracy of the temperature calibration and determine corrective temperature offsets.

Keywords: Hydrodynamics; Sedimentation velocity; Temperature calibration.

Published by Elsevier Inc.

Publication types

Research Support, N.I.H., Intramural

MeSH terms

Calibration
Hydrodynamics
Proteins / analysis
Temperature
Ultracentrifugation* / standards

Substances

Proteins

Grants and funding

ZIA EB000051-06/Intramural NIH HHS/United States