What to do if the protein bands in Western Blot are blurred?

1. The blurred band may be that your protein signal is poor, there is protein degradation or low protein expression in the samples. This can be solved by concentrating the purified protein sample. It is also possible that there is a problem with the antibody, has your antibody been recycled many times? If there are no problems with the sample, you can incubate it with a new antibody diluent, which will enhance your protein signal to some extent. At the same time, it is necessary to pay attention to the moderate washing and sealing of the film to avoid a long time to reduce the signal.  

2. The band color is shallow in the middle and deep on both sides may be caused by running gel or clamping too tightly when transferring the film, resulting in uneven gel or band deformation. It is also possible that your sample has insoluble impurities (e.g. colloids) right in the middle of the band, which can be slightly centrifuged before dosing and then supernatant.  

Hope this helps!