May 09, 2024

Public workspaceNative Nanobleach of native nanodiscs

DOI
dx.doi.org/10.17504/protocols.io.ewov19nbklr2/v1
  • 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;
  • 4Department of Pharmacology, Yale University, New Haven, CT, USA
Caroline Brown
Open access
DOI: dx.doi.org/10.17504/protocols.io.ewov19nbklr2/v1
Protocol Citation: Caroline Brown, Gerard Walker, Snehasish Ghosh, Kallol Gupta, Moitrayee Bhattacharyya 2024. Native Nanobleach of native nanodiscs. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov19nbklr2/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 09, 2024
Last Modified: May 09, 2024
Protocol Integer ID: 99521
Abstract
This is a protocol to conduct Native Nanobleach of native nanodiscs.
Note, this method is adapted from Walker, Brown, et al. (DOI: 10.1038/s41565-023-01547-4)
Prepare flow chamber by treating with poly-L-lysine (PLL) PEG and PEG-biotin.
Add streptavidin to flow chamber.
Add biotinylated GFP-nanobody to flow chamber.
Add native nanodiscs to the flow chamber for immobilization.
Conduct step-photobleaching analysis using TIRF microscopy.