小中大【求助】2D严重的横条纹
菌体样品处理: 1.液氮研磨成粉状; 2.于7M Urea, 2M thiourea, 4% CHAPS, 40mM DTT, 0.5%IPG buffer涡旋1h; 3.离心取上清; 4.加入4倍体积的ice-cold acetone(40mM DTT), -20C过夜,并用ice-cold acetone洗两次; 5于7M Urea, 2M thiourea, 4% CHAPS, 40mM DTT, 0.5%IPG buffer涡旋1h; 6 Broad法蛋白定量.
上样量为300ug,18cm 3-10AMSHAM胶条,70kvh
蛋白集中于酸性端而没有分开,请各位高手指教?