I am doing research on seed storage protein profiling through SDS-PAGE of lentil (Lens culinaris Medik.).But I am facing with a great problem, I am not getting good bands when i run crude extract of protein. i tried with different temperature for boiling extract prior to load on Gel ( at 50, 56, 100 and so on for 5 min or 3 min or 2 min). Everyday I getting new new problems. For extraction I used Tris Hcl buffer, for staining Comassie blue stain. can you tell me what will be the problem and an appropriate suggestion?