Western blot figures for publication: the rules
Western blot figures are the most scrutinized data for image manipulation. The rules boil down to one principle: never change or hide information present in the original.
Dividing lines are added automatically when you reorder lanes, and the uncropped original with quantified regions goes into the slides.
Open the western blot tool →1. Acceptable adjustments
- Apply brightness/contrast uniformly to the whole image, never to individual bands.
- Adjustments must not make bands disappear or appear.
- Disclose non-linear (gamma) adjustments in the figure legend.
2. Rearranging or removing lanes
If you rearrange lanes or remove lanes from the middle of one blot, add a vertical dividing line at each join and explain it in the legend. Presenting different blots as one image is not allowed.
3. What the figure must show
- Molecular-weight markers (kDa) next to the bands
- Lane labels — conditions as a +/− table or names
- Loading control, ideally from the same membrane (state it if not)
- Mention separate exposures in the legend
4. Submitting uncropped originals
Many journals require uncropped images of every blot shown, as supplementary data, with markers visible and the cropped regions indicated. Organize originals by date and condition from day one.
5. Common reviewer flags
- Bands or background look duplicated
- Lanes spliced without dividing lines
- A loading control reused from another experiment
- Cropped so tightly that molecular weight cannot be judged
- Saturated bands used for quantification
Dividing lines are added automatically when you reorder lanes, and the uncropped original with quantified regions goes into the slides.
Open the western blot tool →