tricine-SDS-PAGE小分子电泳体系
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3.1.3.4 Tricine-SDS-PAGE小分子电泳体系
(1)电泳缓冲液的配制
①正极缓冲液anode buffer(1×):0.2M Tris,pH 8.9;
②负极缓冲液cathode buffer(1×):0.1M Tris,0.1M Tricine,0.1% W/V SDS;
③凝胶缓冲液gel buffer(G-B, 3×):3M Tris,0.3%W/V SDS,pH
8.45;
(2)电泳凝胶储存液的配制
①AB-3 储存液(49.5%T, 3%C):0.48g/mL Acrylamide,0.015 g/mL Bis- acrylamide;
(3)还原性loading buffer(4×)
0.15M Tris-HCl(pH 6.8),12%w/v SDS,30%v/v甘油(glycerol),600mM DTT,0.05% 溴酚蓝;
(4)非还原性loading buffer(2×)
0.125 mol/L三羟甲基氨基甲烷盐酸(Tris-HC1),pH 6.8;4%SDS(w/v),20%甘油(v/v),0.02%溴酚蓝(w/v)
(5)固定液(fixing solution)
50%(V/V)甲醇(可用乙醇替代),10%(V/V)冰乙酸,0.1M醋酸铵(Ammonium acetate);
(6)考马斯亮蓝G-250染色液(staining solution)
0.025%(W/V)考马斯亮蓝G-250,10%(V/V)冰乙酸;
(7)脱色液(destaining solution)10%(V/V)冰乙酸;