PT  - JOURNAL ARTICLE
AU  - Tino Pleiner
AU  - Mark Bates
AU  - Dirk Görlich
TI  - A toolbox of anti-mouse and rabbit IgG secondary nanobodies
AID  - 10.1101/209742
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 209742
4099  - http://biorxiv.org/content/early/2017/10/26/209742.short
4100  - http://biorxiv.org/content/early/2017/10/26/209742.full
AB  - Polyclonal 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 E. 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, both in peroxidase- and fluorophore-linked form. Their site-specific labeling with multiple fluorophores creates bright imaging reagents for confocal and super-resolution microscopy with much smaller label displacement than traditional secondary antibodies. They also enable simpler and faster immunostaining protocols and even allow multi-target localization with primary IgGs from the same species and of the same class.