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@ARTICLE{Simons:860092,
author = {Simons, Indra M. and Mohrlüder, Jeannine and Feederle,
Regina and Kremmer, Elisabeth and Zobel, Thomas and Dobner,
Jochen and Bleffert, Nicole and Hoffmann, Silke and
Willbold, Dieter},
title = {{T}he highly {GABARAP} specific rat monoclonal antibody
8{H}5 visualizes {GABARAP} in immunofluorescence imaging at
endogenous levels},
journal = {Scientific reports},
volume = {9},
number = {1},
issn = {2045-2322},
address = {[London]},
publisher = {Macmillan Publishers Limited, part of Springer Nature},
reportid = {FZJ-2019-00878},
pages = {526},
year = {2019},
abstract = {The determination of unique functions of GABARAP
(gamma-aminobutyric acid type A receptor-associated
protein), a member of the highly conserved protein family of
mammalian autophagy-related 8 protein (mATG8), within
diverse cellular processes remains challenging. Because
available anti-GABARAP antibodies perform inadequate,
especially within various microscopy-based applications, we
aimed to develop an antibody that targets GABARAP but not
its close orthologs. Following the latest recommendations
for antibody validation including fluorescence protein
tagging, genetic and orthogonal strategies, we characterized
the resulting anti-GABARAP (8H5) antibody during confocal
immunofluorescence imaging in-depth. We compared the
antibody staining pattern with that obtained for
fluorescence protein tagged GABARAP, GABARAPL1 or GABARAPL2
each ectopically expressed in GABARAP knockout cells.
Furthermore, we imaged cells expressing all mATG8 family
members at endogenous levels and checked GABARAP knockout
cells for unspecific staining under fed or
macroautophagy-inducing conditions. Finally, we
simultaneously stained cells for endogenous GABARAP and the
common autophagosomal marker LC3B. Summarized, the presented
antibody shows high specificity for GABARAP without
cross-reactivity to other mATG8 family members in
immunofluorescence imaging making it a valuable tool for the
identification of unique GABARAP functions.},
cin = {ICS-6},
ddc = {600},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {552 - Engineering Cell Function (POF3-552)},
pid = {G:(DE-HGF)POF3-552},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30679523},
UT = {WOS:000456553400091},
doi = {10.1038/s41598-018-36717-1},
url = {https://juser.fz-juelich.de/record/860092},
}