Cargando…
Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them
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 gro...
Autores principales: | , , , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9518894/ https://www.ncbi.nlm.nih.gov/pubmed/36095010 http://dx.doi.org/10.1371/journal.pbio.3001783 |
_version_ | 1784799285508833280 |
---|---|
author | Kroon, Cristina Breuer, Larissa Jones, Lydia An, Jeehye Akan, Ayça Mohamed Ali, Elkhansa Ahmed Busch, Felix Fislage, Marinus Ghosh, Biswajit Hellrigel-Holderbaum, Max Kazezian, Vartan Koppold, Alina Moreira Restrepo, Cesar Alberto Riedel, Nico Scherschinski, Lea Urrutia Gonzalez, Fernando Raúl Weissgerber, Tracey L. |
author_facet | Kroon, Cristina Breuer, Larissa Jones, Lydia An, Jeehye Akan, Ayça Mohamed Ali, Elkhansa Ahmed Busch, Felix Fislage, Marinus Ghosh, Biswajit Hellrigel-Holderbaum, Max Kazezian, Vartan Koppold, Alina Moreira Restrepo, Cesar Alberto Riedel, Nico Scherschinski, Lea Urrutia Gonzalez, Fernando Raúl Weissgerber, Tracey L. |
author_sort | Kroon, Cristina |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9518894 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-95188942022-09-29 Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them Kroon, Cristina Breuer, Larissa Jones, Lydia An, Jeehye Akan, Ayça Mohamed Ali, Elkhansa Ahmed Busch, Felix Fislage, Marinus Ghosh, Biswajit Hellrigel-Holderbaum, Max Kazezian, Vartan Koppold, Alina Moreira Restrepo, Cesar Alberto Riedel, Nico Scherschinski, Lea Urrutia Gonzalez, Fernando Raúl Weissgerber, Tracey L. PLoS Biol Meta-Research Article 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. Public Library of Science 2022-09-12 /pmc/articles/PMC9518894/ /pubmed/36095010 http://dx.doi.org/10.1371/journal.pbio.3001783 Text en © 2022 Kroon 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 | Meta-Research Article Kroon, Cristina Breuer, Larissa Jones, Lydia An, Jeehye Akan, Ayça Mohamed Ali, Elkhansa Ahmed Busch, Felix Fislage, Marinus Ghosh, Biswajit Hellrigel-Holderbaum, Max Kazezian, Vartan Koppold, Alina Moreira Restrepo, Cesar Alberto Riedel, Nico Scherschinski, Lea Urrutia Gonzalez, Fernando Raúl Weissgerber, Tracey L. Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
title | Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
title_full | Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
title_fullStr | Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
title_full_unstemmed | Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
title_short | Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
title_sort | blind spots on western blots: assessment of common problems in western blot figures and methods reporting with recommendations to improve them |
topic | Meta-Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9518894/ https://www.ncbi.nlm.nih.gov/pubmed/36095010 http://dx.doi.org/10.1371/journal.pbio.3001783 |
work_keys_str_mv | AT krooncristina blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT breuerlarissa blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT joneslydia blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT anjeehye blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT akanayca blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT mohamedalielkhansaahmed blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT buschfelix blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT fislagemarinus blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT ghoshbiswajit blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT hellrigelholderbaummax blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT kazezianvartan blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT koppoldalina blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT moreirarestrepocesaralberto blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT riedelnico blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT scherschinskilea blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT urrutiagonzalezfernandoraul blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem AT weissgerbertraceyl blindspotsonwesternblotsassessmentofcommonproblemsinwesternblotfiguresandmethodsreportingwithrecommendationstoimprovethem |