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Summary
| Description |
English: Western blotting is a standard laboratory method that uses antibodies to detect target proteins in a sample. (1) The sample, typically a mixture of proteins, is loaded on the gel. A molecular weight (MW) marker, which contains prelabeled proteins of varied, known molecular weights, is loaded on the gel alongside the protein sample as a size reference. (2) Gel electrophoresis is used to separate proteins based on their molecular weight. (3) The proteins are transferred, or “blotted”, onto a membrane. (4) The membrane is blocked to reduce nonspecific binding and then sequentially probed with a primary antibody that specifically binds to the protein of interest and a secondary antibody. The latter binds the primary antibody and carries an enzyme or a fluorophore that allows subsequent detection. (5) The signal is detected through a chemiluminescent reaction or fluorescence, respectively. (6) An image of the western blot is prepared for publication: Annotations are added and often the blot is cropped. |
| Date | |
| Source | Kroon C, Breuer L, Jones L, An J, Akan A, Mohamed Ali EA, et al. (2022) Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them. PLoS Biol 20(9): e3001783. https://doi.org/10.1371/journal.pbio.3001783 |
| Author | 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 |
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| Date/Time | Thumbnail | Dimensions | User | Comment | |
|---|---|---|---|---|---|
| current | 01:43, 15 December 2022 |  | 1,500 × 1,499 (558 KB) | Guest2625 | Uploaded a work 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 from Kroon C, Breuer L, Jones L, An J, Akan A, Mohamed Ali EA, et al. (2022) Blind spots on western blots: Assessment of common problems in western blot figures... |
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