Decrease or titrate downwards the amount of protein you put in each lane. Also, try to always make use of new APS and TEMED so as to get a clear gel. I think that should help.

Thank you very much

From this picture, it's clear that your protein are not separating. For me, you should check first the procedure of preparation of your samples. The reagents you are using, teh conditions especially pay attention to the steps that can influenc the temperature or the pH of the samples. for western blot, checking the buffers is very important, but what I can suggest is that you increase the concentration of your gel, so you can seperate your proteins better. finally, to avoid non-specifi bands, blocking step is very important to minimize them.

you probably consider some details:

1.- Protein concentration and integrity

2.- PBS-tween or TBS-Tween buffer to use

3.- Ab´s primary and secondary dilution is important

4.- Blocking conditions: Dry milk or BSA from 5 to 15%

the concentration of gel will depend on the size of the detected band!