A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA).
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2022
2022
Historique:
received:
06
05
2022
accepted:
20
05
2022
entrez:
1
7
2022
pubmed:
2
7
2022
medline:
8
7
2022
Statut:
epublish
Résumé
The Rho family Guanine nucleotide exchange factor (GEF) ARHGEF17 (also known as TEM4) is a large protein with only 3 annotated regions: an N-terminal actin-binding domain, a Rho-specific dbl homology (DH)- pleckstrin homology (PH) type GEF domain and a seven bladed β propeller fold at the C-terminus with unknown function. TEM4 has been implicated in numerous activities that rely on regulation of the cytoskeleton including cell migration, cell-cell junction formation and the spindle assembly checkpoint during mitosis. Here we have assessed the specificity of a TEM4 polyclonal antibody that has been commonly used as a Western blotting and immunocytochemistry probe for TEM4 in mammalian cells. We find that this antibody, in addition to its intended target, cross-reacts with the Nuclear Mitotic Apparatus Protein 1 (NuMA) in Western blotting and immunoprecipitation, and detects NuMA preferentially in immunocytochemistry. This cross-reactivity, with an abundant chromatin- and mitotic spindle-associated factor, is likely to affect the interpretation of experiments that make use of this antibody probe, in particular by immunocytochemistry and immunoprecipitation.
Identifiants
pubmed: 35776709
doi: 10.1371/journal.pone.0268848
pii: PONE-D-22-13298
pmc: PMC9249204
doi:
Substances chimiques
Actins
0
Antibodies
0
Cell Cycle Proteins
0
Rho Guanine Nucleotide Exchange Factors
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0268848Subventions
Organisme : CIHR
ID : 376557
Pays : Canada
Organisme : CIHR
ID : 388292
Pays : Canada
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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