Fluorescence size exclusion chromatography (FSEC) of native nanodiscs

Caroline Brown, Snehasish Ghosh, Kallol Gupta

Published: 2024-05-10 DOI: 10.17504/protocols.io.14egn676ml5d/v1

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Abstract

This is a protocol describing how to conduct Fluorescence size exclusion chromatography of native nanodiscs.

Steps

1.

Conduct extraction into native nanodiscs following established protocols.

2.

Inject 200uL native nanodiscs onto a Superose6 (10/300GL) column which is pre-equilibrated with TBS.

3.

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.

4.

Collect eluent for downstream analysis.

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