Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them.

Autor: Kroon C; Institute of Molecular Biology and Biochemistry, Charité-Universitätsmedizin Berlin, Berlin, Germany., Breuer L; German Center for Neurodegenerative Diseases (DZNE) within the Helmholtz Association, Berlin, Germany.; Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, Berlin, Germany., Jones L; Berlin School of Public Health, Charité-Universitätsmedizin Berlin, Berlin, Germany., An J; Berlin School of Mind and Brain, Humboldt-Universität zu Berlin, Berlin, Germany.; Department of Experimental Neurology and Center for Stroke Research, Charité-Universitätsmedizin Berlin, Berlin, Germany., Akan A; Berlin School of Mind and Brain, Humboldt-Universität zu Berlin, Berlin, Germany.; Max Planck Institute for Human Cognitive and Brain Sciences, Leipzig, Germany., Mohamed Ali EA; Department of Psychiatry and Neurosciences, Charité-Universitätsmedizin Berlin, Berlin, Germany., Busch F; Department of Radiology, Charité-Universitätsmedizin Berlin, Berlin, Germany.; Department of Anesthesiology and Intensive Care Medicine, Charité-Universitätsmedizin Berlin, Berlin, Germany., Fislage M; Department of Anesthesiology and Intensive Care Medicine, Charité-Universitätsmedizin Berlin, Berlin, Germany., Ghosh B; Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, Berlin, Germany.; Max Planck Institute for Molecular Genetics, Berlin, Germany., Hellrigel-Holderbaum M; Berlin School of Mind and Brain, Humboldt-Universität zu Berlin, Berlin, Germany.; Interdisciplinary Center of Sleep Medicine, Charité-Universitätsmedizin Berlin, Berlin, Germany., Kazezian V; QUEST Center for Responsible Research, Berlin Institute of Health at Charité-Universitätsmedizin Berlin, Berlin, Germany.; Department of Biochemistry, Freie Universität Berlin, Berlin, Germany., Koppold A; Institute for Systems Neuroscience, University Medical Center Hamburg-Eppendorf, Hamburg, Germany., Moreira Restrepo CA; Berlin School of Public Health, Charité-Universitätsmedizin Berlin, Berlin, Germany., Riedel N; QUEST Center for Responsible Research, Berlin Institute of Health at Charité-Universitätsmedizin Berlin, Berlin, Germany., Scherschinski L; Department of Neurosurgery, Charité-Universitätsmedizin Berlin, Berlin, Germany., Urrutia Gonzalez FR; Berlin School of Public Health, Charité-Universitätsmedizin Berlin, Berlin, Germany.; Institute of Public Health, Charité-Universitätsmedizin Berlin, Berlin, Germany.; Center for Stroke Research Berlin, Charité-Universitätsmedizin Berlin, Berlin, Germany., Weissgerber TL; QUEST Center for Responsible Research, Berlin Institute of Health at Charité-Universitätsmedizin Berlin, Berlin, Germany.
Jazyk: angličtina
Zdroj: PLoS biology [PLoS Biol] 2022 Sep 12; Vol. 20 (9), pp. e3001783. Date of Electronic Publication: 2022 Sep 12 (Print Publication: 2022).
DOI: 10.1371/journal.pbio.3001783
Abstrakt: Western blotting is a standard laboratory method used to detect proteins and assess their expression levels. Unfortunately, poor western blot image display practices and a lack of detailed methods reporting can limit a reader's ability to evaluate or reproduce western blot results. While several groups have studied the prevalence of image manipulation or provided recommendations for improving western blotting, data on the prevalence of common publication practices are scarce. We systematically examined 551 articles published in the top 25% of journals in neurosciences (n = 151) and cell biology (n = 400) that contained western blot images, focusing on practices that may omit important information. Our data show that most published western blots are cropped and blot source data are not made available to readers in the supplement. Publishing blots with visible molecular weight markers is rare, and many blots additionally lack molecular weight labels. Western blot methods sections often lack information on the amount of protein loaded on the gel, blocking steps, and antibody labeling protocol. Important antibody identifiers like company or supplier, catalog number, or RRID were omitted frequently for primary antibodies and regularly for secondary antibodies. We present detailed descriptions and visual examples to help scientists, peer reviewers, and editors to publish more informative western blot figures and methods. Additional resources include a toolbox to help scientists produce more reproducible western blot data, teaching slides in English and Spanish, and an antibody reporting template.
Competing Interests: The authors have declared the no competing interests exist.
Databáze: MEDLINE
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