og:image: http://jcb.rupress.org/sites/default/files/highwire/jcb/217/3.cover-source.jpg citation_mjid: jcb;217/3/1143 article:published_time: 2018-03-05 og:site_name: JCB citation_reference: citation_journal_title=FEBS Letters;citation_journal_abbrev=FEBS Letters;citation_author=M. Arbabi Ghahroudi;citation_author=A. Desmyter;citation_author=L. Wyns;citation_author=R. Hamers;citation_author=S. Muyldermans;citation_title=Selection and identification of single domain antibody fragments from camel heavy-chain antibodies.;citation_pages=521-526;citation_volume=414;citation_year=1997;citation_issue=3;citation_pmid=9323027;citation_doi=10.1016/S0014-5793(97)01062-4 citation_journal_title: J Cell Biol type: article og:description: 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. citation_author_email: tino.pleiner@mpibpc.mpg.de citation_issn: 0021-9525 citation_full_html_url: http://jcb.rupress.org/content/217/3/1143.full citation_public_url: http://jcb.rupress.org/content/217/3/1143 dc:title: A toolbox of anti?mouse and anti?rabbit IgG secondary nanobodies | JCB Content-Encoding: UTF-8 citation_pdf_url: http://jcb.rupress.org/content/217/3/1143.full.pdf citation_section: Research Article citation_lastpage: 1154 citation_fulltext_world_readable: citation_journal_abbrev: J Cell Biol DC.Identifier: 10.1083/jcb.201709115 DC.Rights: © 2018 Pleiner et al.. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). citation_author: Tino Pleiner citation_abstract_html_url: http://jcb.rupress.org/content/217/3/1143.abstract citation_issue: 3 HW.identifier: /jcb/217/3/1143.atom citation_doi: 10.1083/jcb.201709115 DC.Relation: 10.1083/jcb.201802025 citation_volume: 217 Content-Language: en Generator: Drupal 7 (http://drupal.org) citation_author_orcid: http://orcid.org/0000-0002-5104-0315 DC.AccessRights: open-access citation_publication_date: 2018/03/05 citation_title: A toolbox of anti?mouse and anti?rabbit IgG secondary nanobodies citation_author_institution: Department of Cellular Logistics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany citation_publisher: Rockefeller University Press citation_id: 217/3/1143 title: A toolbox of anti?mouse and anti?rabbit IgG secondary nanobodies | JCB DC.Description: 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. Content-Type-Hint: text/html; charset=utf-8 DC.Format: text/html DC.Publisher: Rockefeller University Press DC.Contributor: Tino Pleiner Content-Type: application/xhtml+xml; charset=UTF-8 X-Parsed-By: org.apache.tika.parser.DefaultParser og:type: article article:section: Research Article citation_pmid: 29263082 citation_article_type: Research Article og:title: A toolbox of anti?mouse and anti?rabbit IgG secondary nanobodies citation_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.

DC.Title: A toolbox of anti?mouse and anti?rabbit IgG secondary nanobodies issue_cover_image: http://jcb.rupress.org/sites/default/files/highwire/jcb/217/3.cover-source.jpg citation_firstpage: 1143 viewport: width=device-width, initial-scale=1, maximum-scale=3, minimum-scale=1, user-scalable=yes citation_funding_source: citation_funder=Max-Planck-Gesellschaft;citation_funder_id=https://doi.org/10.13039/501100004189; HW.pisa: jcb;217/3/1143 DC.Language: en DC.Date: 2018-03-05 citation_access: all category: research-article og:url: http://jcb.rupress.org/content/217/3/1143 article_thumbnail: http://jcb.rupress.org/content/jcb/217/3/1143/embed/icon-1.gif