Native Nanobleach of native nanodiscs
Caroline Brown, Snehasish Ghosh, Kallol Gupta, Gerard Walker, Moitrayee Bhattacharyya
Published: 2024-05-10 DOI: 10.17504/protocols.io.ewov19nbklr2/v1
Abstract
This is a protocol to conduct Native Nanobleach of native nanodiscs.
Steps
1.
Note, this method is adapted from Walker, Brown, et al. (DOI: 10.1038/s41565-023-01547-4) 10.1038/s41565-023-01547-4)
2.
Prepare flow chamber by treating with poly-L-lysine (PLL) PEG and PEG-biotin.
3.
Add streptavidin to flow chamber.
4.
Add biotinylated GFP-nanobody to flow chamber.
5.
Add native nanodiscs to the flow chamber for immobilization.
6.
Conduct step-photobleaching analysis using TIRF microscopy.
