Confirming Antibody Specificity in Xenopus
- 1Molecular Embryology Laboratory, University of Portsmouth, Portsmouth, Hampshire PO1 2DY, United Kingdom;
- 2European Xenopus Resource Centre, Institute of Biological and Biomedical Sciences, University of Portsmouth, Portsmouth, Hampshire PO1 2DY, United Kingdom
- ↵ 3Correspondence: matthew.guille{at}port.ac.uk
Abstract
Verifying that a new antibody recognizes its target can be difficult. In this protocol, expression of a target protein in Xenopus embryos is either knocked down using CRISPR–Cas9 technology (for zygotic proteins) or enhanced by microinjection of a synthetic mRNA (for maternal proteins). Western blotting analysis is then performed. If the antibody recognizes the target protein, the western blot will show a relatively weak band for CRISPR-injected embryos and a relatively strong band for RNA-injected embryos. This represents a straightforward, powerful strategy for confirming antibody specificity in Xenopus.
Footnotes
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From the Xenopus collection, edited by Hazel L. Sive.
