Hi!! Someone know best method to analyze SUMO1 by western blot? I tried different buffers to extract proteins, different loading buffers, different antibodies and doesnt work :S:S I'm getting mad really, it shouldnt be too difficoult
Thanks so much Bruna, yes I used RIPA buffer suplied with NEM (N-Ethylmaleimide), a specific inhibitor of sumo protease, I used RIPA without it, I used mPER, I used a buffer with urea, I used laemli like loading buffer, I used other loading buffer, I did acrilamide gel with SDS, I used the ab from abcam and I used other ab from CST... and really i dont know what more to do, it doesnt work...what is your positive control?? I want detect endogenous sumo, is not an overexpression.