Cargando…

A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Prifti, Diogjena Katerina, Lauzier, Annie, Elowe, Sabine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9249204/
https://www.ncbi.nlm.nih.gov/pubmed/35776709
http://dx.doi.org/10.1371/journal.pone.0268848
_version_ 1784739526464241664
author Prifti, Diogjena Katerina
Lauzier, Annie
Elowe, Sabine
author_facet Prifti, Diogjena Katerina
Lauzier, Annie
Elowe, Sabine
author_sort Prifti, Diogjena Katerina
collection PubMed
description 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.
format Online
Article
Text
id pubmed-9249204
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-92492042022-07-02 A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA) Prifti, Diogjena Katerina Lauzier, Annie Elowe, Sabine PLoS One Research Article 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. Public Library of Science 2022-07-01 /pmc/articles/PMC9249204/ /pubmed/35776709 http://dx.doi.org/10.1371/journal.pone.0268848 Text en © 2022 Prifti et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Prifti, Diogjena Katerina
Lauzier, Annie
Elowe, Sabine
A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)
title A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)
title_full A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)
title_fullStr A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)
title_full_unstemmed A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)
title_short A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA)
title_sort commercial arhgef17/tem4 antibody cross-reacts with nuclear mitotic apparatus protein 1 (numa)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9249204/
https://www.ncbi.nlm.nih.gov/pubmed/35776709
http://dx.doi.org/10.1371/journal.pone.0268848
work_keys_str_mv AT priftidiogjenakaterina acommercialarhgef17tem4antibodycrossreactswithnuclearmitoticapparatusprotein1numa
AT lauzierannie acommercialarhgef17tem4antibodycrossreactswithnuclearmitoticapparatusprotein1numa
AT elowesabine acommercialarhgef17tem4antibodycrossreactswithnuclearmitoticapparatusprotein1numa
AT priftidiogjenakaterina commercialarhgef17tem4antibodycrossreactswithnuclearmitoticapparatusprotein1numa
AT lauzierannie commercialarhgef17tem4antibodycrossreactswithnuclearmitoticapparatusprotein1numa
AT elowesabine commercialarhgef17tem4antibodycrossreactswithnuclearmitoticapparatusprotein1numa