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The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence

RNA-binding protein Fused-in Sarcoma (FUS) plays an essential role in various cellular processes. Mutations in the C-terminal domain region, where the nuclear localization signal (NLS) is located, causes the redistribution of FUS from the nucleus to the cytoplasm. In neurons, neurotoxic aggregates a...

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Autores principales: Alshalfie, Walaa, Fotouhi, Maryam, Ayoubi, Riham, You, Zhipeng, Southern, Kathleen, McPherson, Peter S., Laflamme, Carl
Formato: Online Artículo Texto
Lenguaje:English
Publicado: F1000 Research Limited 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10293799/
https://www.ncbi.nlm.nih.gov/pubmed/37384305
http://dx.doi.org/10.12688/f1000research.133220.2
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author Alshalfie, Walaa
Fotouhi, Maryam
Ayoubi, Riham
You, Zhipeng
Southern, Kathleen
McPherson, Peter S.
Laflamme, Carl
author_facet Alshalfie, Walaa
Fotouhi, Maryam
Ayoubi, Riham
You, Zhipeng
Southern, Kathleen
McPherson, Peter S.
Laflamme, Carl
author_sort Alshalfie, Walaa
collection PubMed
description RNA-binding protein Fused-in Sarcoma (FUS) plays an essential role in various cellular processes. Mutations in the C-terminal domain region, where the nuclear localization signal (NLS) is located, causes the redistribution of FUS from the nucleus to the cytoplasm. In neurons, neurotoxic aggregates are formed as a result, contributing to neurogenerative diseases. Well-characterized anti-FUS antibodies would enable the reproducibility of FUS research, thereby benefiting the scientific community.  In this study, we characterized ten FUS commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.
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spelling pubmed-102937992023-06-28 The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence Alshalfie, Walaa Fotouhi, Maryam Ayoubi, Riham You, Zhipeng Southern, Kathleen McPherson, Peter S. Laflamme, Carl F1000Res Data Note RNA-binding protein Fused-in Sarcoma (FUS) plays an essential role in various cellular processes. Mutations in the C-terminal domain region, where the nuclear localization signal (NLS) is located, causes the redistribution of FUS from the nucleus to the cytoplasm. In neurons, neurotoxic aggregates are formed as a result, contributing to neurogenerative diseases. Well-characterized anti-FUS antibodies would enable the reproducibility of FUS research, thereby benefiting the scientific community.  In this study, we characterized ten FUS commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs. F1000 Research Limited 2023-06-26 /pmc/articles/PMC10293799/ /pubmed/37384305 http://dx.doi.org/10.12688/f1000research.133220.2 Text en Copyright: © 2023 Alshalfie W et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Data Note
Alshalfie, Walaa
Fotouhi, Maryam
Ayoubi, Riham
You, Zhipeng
Southern, Kathleen
McPherson, Peter S.
Laflamme, Carl
The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence
title The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence
title_full The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence
title_fullStr The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence
title_full_unstemmed The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence
title_short The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence
title_sort identification of high-performing antibodies for rna-binding protein fus for use in western blot, immunoprecipitation, and immunofluorescence
topic Data Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10293799/
https://www.ncbi.nlm.nih.gov/pubmed/37384305
http://dx.doi.org/10.12688/f1000research.133220.2
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