XB-ART-57444
Cold Spring Harb Protoc
2020 Dec 01;202012:. doi: 10.1101/pdb.prot105601.
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Confirming Antibody Specificity in Xenopus.
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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.
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Species referenced: Xenopus laevis