A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies
| العنوان: | A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies |
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| المؤلفون: | Mark Bates, Tino Pleiner, Dirk Görlich |
| المصدر: | The Journal of Cell Biology Journal of Cell Biology |
| بيانات النشر: | The Rockefeller University Press, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Animal Use Alternatives, Fluorophore, Indoles, Blotting, Western, Antibody Affinity, medicine.disease_cause, law.invention, Tools, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, law, Antibody Specificity, Commentaries, medicine, Escherichia coli, Animals, Spotlight, Research Articles, biology, Optical Imaging, Antibodies, Monoclonal, Cell Biology, Single-Domain Antibodies, Primary and secondary antibodies, Molecular biology, Recombinant Proteins, Blot, 030104 developmental biology, chemistry, Biochemistry, Polyclonal antibodies, Immunoglobulin G, Recombinant DNA, biology.protein, Female, Rabbits, Antibody, Camelids, New World, 030217 neurology & neurosurgery, Immunostaining |
| الوصف: | Pleiner, Bates, and Görlich introduce anti–mouse and anti–rabbit IgG nanobodies that can be produced in E. coli and fused to reporters or labeled fluorescently to create bright and specific detection reagents with unique advantages over conventional polyclonal secondary antibodies. 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. |
| وصف الملف: | application/pdf; application/vnd.ms-excel; application/xhtml+xml |
| اللغة: | English |
| تدمد: | 1540-8140 0021-9525 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b5bb2cbf08b66635a6fc9d35834a00e7 http://europepmc.org/articles/PMC5839796 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....b5bb2cbf08b66635a6fc9d35834a00e7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15408140 00219525 |
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