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

Authors :
Alshalfie W
Fotouhi M
Ayoubi R
You Z
Southern K
McPherson PS
Laflamme C
Source :
F1000Research [F1000Res] 2023 Jun 26; Vol. 12, pp. 376. Date of Electronic Publication: 2023 Jun 26 (Print Publication: 2023).
Publication Year :
2023

Abstract

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.<br />Competing Interests: Competing interests: For this project, the laboratory of Peter McPherson developed partnerships with high-quality antibody manufacturers and knockout cell line providers. The partners provide antibodies and knockout cell lines to the McPherson laboratory at no cost. These partners include: - Abcam- ABclonal -Aviva Systems Biology -Bio Techne -Cell Signalling Technology -Developmental Studies Hybridoma Bank -GeneTex – Horizon Discovery – Proteintech – Synaptic Systems -Thermo Fisher Scientific.<br /> (Copyright: © 2023 Alshalfie W et al.)

Details

Language :
English
ISSN :
2046-1402
Volume :
12
Database :
MEDLINE
Journal :
F1000Research
Publication Type :
Academic Journal
Accession number :
37384305
Full Text :
https://doi.org/10.12688/f1000research.133220.2