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

by Cristina Kroon, Larissa Breuer, Lydia Jones, Jeehye An, Ayça Akan, Elkhansa Ahmed Mohamed Ali, Felix Busch, Marinus Fislage, Biswajit Ghosh, Max Hellrigel-Holderbaum, Vartan Kazezian, Alina Koppold, Cesar Alberto Moreira Restrepo, Nico Riedel, Lea Scherschinski, Fernando Raúl Urrutia Gonzalez, Tracey L. Weissgerber

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.