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@ARTICLE{Tan:1009340,
author = {Tan, Hua and Bungert-Plümke, Stefanie and Kortzak, Daniel
and Fahlke, Christoph and Stölting, Gabriel},
title = {{D}ual-color {C}olocalization in {S}ingle-molecule
{L}ocalization {M}icroscopy to {D}etermine the {O}ligomeric
{S}tate of {P}roteins in the {P}lasma {M}embrane},
journal = {Bio-protocol},
volume = {13},
number = {13},
issn = {2331-8325},
address = {Sunnyvale, CA},
publisher = {bio-protocol.org},
reportid = {FZJ-2023-02769},
pages = {4749},
year = {2023},
note = {Development of this protocol was supported by the Deutsche
Forschungsgemeinschaft (FA 301/15-1 to Ch.F.) as part of
Research Unit FOR 5046, project P4. We acknowledge the
original research paper (Tan et al., 2022), from which this
protocol is derived, as the primary reference when
additional information is needed.},
abstract = {Determining the oligomeric state of membrane proteins is
critical for understanding their function. However,
traditional ex situ methods like clear native gel
electrophoresis can disrupt protein subunit interactions
during sample preparation. In situ methods such as stepwise
photobleaching have limitations due to high expression
levels and limitations of optical resolution in microscopy.
Super-resolution microscopy techniques such as
single-molecule localization microscopy (SMLM) have the
potential to overcome these limitations, but the stochastic
nature of signals can lead to miscounting due to
over-expression, background noise, and temporal separation
of signals. Additionally, this technique has limited
application due to the limited selection of fluorescent
labels and the demanding control of laser power. To address
these issues, we developed a dual color colocalization (DCC)
strategy that offers higher tolerance to background noise
and simplifies data acquisition and processing for
high-throughput and reliable counting. The DCC strategy was
used to determine the oligomeric states of membrane proteins
of the SLC17 and SLC26 family with SMLM, providing a robust
and efficient method for studying protein interactions.},
cin = {IBI-1},
ddc = {570},
cid = {I:(DE-Juel1)IBI-1-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {37456335},
UT = {WOS:001089325800011},
doi = {10.21769/BioProtoc.4749},
url = {https://juser.fz-juelich.de/record/1009340},
}