To briefly explain my experiment, I add a chemical probe to the protein lysate, the probe covalently binds to several proteins in the lysate and further the probe is tagged to a fluorophore by CLICK chemistry.

I am particularly interested in a protein band at approx 26 kDa, so I run an 18% SDS-PAGE gel to obtain a good resolution of bands. The problem is that the bands appear to be blurry (or diffused) in the fluorescence image(image attached) but seem fine in the coomassie. So, are there any conditions that you follow in your lab to run a high-percentage gel? It would really be helpful. [Ignore the intense bands at the top of the gel, I had to expose it for a longer time to get a band profile of protein bands at lower molecular weights as they had lesser intensity]

Thanks in advance for your answers/recommendations.