May 09, 2024
Fluorescence size exclusion chromatography (FSEC) of native nanodiscs
- Caroline Brown1,2,3,
- Snehasish Ghosh1,2,3,
- Kallol Gupta1,2,3
- 1Nanobiology Institute, Yale University, West Haven, CT, USA;
- 2Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA;
- 3Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815
Protocol Citation: Caroline Brown, Snehasish Ghosh, Kallol Gupta 2024. Fluorescence size exclusion chromatography (FSEC) of native nanodiscs . protocols.io https://dx.doi.org/10.17504/protocols.io.14egn676ml5d/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 04, 2024
Last Modified: May 09, 2024
Protocol Integer ID: 99251
Abstract
This is a protocol describing how to conduct Fluorescence size exclusion chromatography of native nanodiscs.
Conduct extraction into native nanodiscs following established protocols.
Inject 200uL native nanodiscs onto a Superose6 (10/300GL) column which is pre-equilibrated with TBS.
Perform separation at a flow rate of 0.5 mL/min and detect eluent with a Shimadzu fluorometer with excitation set to 488 nm and emission set to 509 nm.
Collect eluent for downstream analysis.