I am trying to dephosphorylate a target protein from a yeast crude extract following a protocol with lambda-phosphatase. In my first try, I finished the reaction by adding 5x loading buffer, then heating and finally SDS-PAGE and blotting. However, I found only aberrant and distorted bands. I imagined the problem was the Brij 35 contained in the buffer. So, I tried again but now I precipitated the sample with TCA, acetone, etc.. but again I got the same results. I wonder if anyone could help me to solve this problem.