Confirming antibody specificity in Xenopus

Research output: Other contribution

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.
Original languageEnglish
TypeProtocol
PublisherCold Spring Harbor Laboratory Press
Number of pages8
Edition12
Volume2020
DOIs
Publication statusPublished - 1 Dec 2020

Publication series

NameCold Spring Harbor Protocols
PublisherCold Spring Harbor Laboratory Press
ISSN (Print)1940-3402

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