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  A toolbox of anti-mouse and anti-rabbit IgG secondary nanobodies.

Pleiner, T., Bates, M., & Görlich, D. (2018). A toolbox of anti-mouse and anti-rabbit IgG secondary nanobodies. Journal of Cell Biology, 217(3), 1143-1154. doi:10.1083/jcb.201709115.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-9AC5-2 Version Permalink: http://hdl.handle.net/21.11116/0000-0001-3F4D-4
Genre: Journal Article

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 Creators:
Pleiner, T.1, Author              
Bates, M.2, Author              
Görlich, D.1, Author              
Affiliations:
1Department of Cellular Logistics, MPI for Biophysical Chemistry, Max Planck Society, ou_578574              
2Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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 Abstract: Polyclonal anti-immunoglobulin G (anti-IgG) secondary antibodies are essential tools for many molecular biology techniques and diagnostic tests. Their animal-based production is, however, a major ethical problem. Here, we introduce a sustainable alternative, namely nanobodies against all mouse IgG subclasses and rabbit IgG. They can be produced at large scale in Escherichia coli and could thus make secondary antibody production in animals obsolete. Their recombinant nature allows fusion with affinity tags or reporter enzymes as well as efficient maleimide chemistry for fluorophore coupling. We demonstrate their superior performance in Western blotting, in both peroxidase- and fluorophore-linked form. Their site-specific labeling with multiple fluorophores creates bright imaging reagents for confocal and superresolution microscopy with much smaller label displacement than traditional secondary antibodies. They also enable simpler and faster immunostaining protocols, and allow multitarget localization with primary IgGs from the same species and of the same class.

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Language(s): eng - English
 Dates: 2017-12-202018-03-05
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1083/jcb.201709115
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Title: Journal of Cell Biology
Source Genre: Journal
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Pages: - Volume / Issue: 217 (3) Sequence Number: - Start / End Page: 1143 - 1154 Identifier: -