Confirming antibody specificity in Xenopus
Research output: Other contribution
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Confirming antibody specificity in Xenopus. / Martin, Sian A.; Page, Suzannah J.; Piccinni, Maya; Guille, Matt.
8 p. 12 ed. Cold Spring Harbor Laboratory Press. 2020, Protocol. (Cold Spring Harbor Protocols).Research output: Other contribution
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TY - GEN
T1 - Confirming antibody specificity in Xenopus
AU - Martin, Sian A.
AU - Page, Suzannah J.
AU - Piccinni, Maya
AU - Guille, Matt
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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.
U2 - 10.1101/pdb.prot105601
DO - 10.1101/pdb.prot105601
M3 - Other contribution
VL - 2020
T3 - Cold Spring Harbor Protocols
PB - Cold Spring Harbor Laboratory Press
ER -
ID: 25019862