Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

EPR spectroscopy

OJ Owen D. Jarman
OB Olivier Biner
JW John J. Wright
JH Judy Hirst
request Request a Protocol
ask Ask a question
Favorite

EPR samples were prepared anaerobically by transferring 100 µL of purified complex I (5.8 mg mL–1) to a 4 mm (O.D.) EPR tube (Wilmad), then the complex I was reduced by the addition of 15 mM NADH. The sample was immediately frozen in dry-ice/acetone before transfer to liquid nitrogen for storage. EPR measurements were performed using an X/Q-band Bruker Elexsys E580 Spectrometer (Bruker BioSpin) operating in X-band mode and equipped with a closed-cycle cryostat (Cryogenic Ltd) and X-band split-ring resonator module (ER 4118X-MD5). EPR measurement conditions were 100 kHz modulation frequency, 7 G modulation amplitude, 2 mW microwave power; other relevant measurement conditions are given in the figure legend. All spectra presented are baseline corrected using a buffer-only sample. EPR simulations were performed using the EasySpin package for MATLAB74.

Copyright and License information: The Author(s) ©2021
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A