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1、SDS-PAGE电泳初级指南SDS-PAGE 电泳14% Resolving gel solution (5ml)30% Acrylamide solution2.33ml4 X resolving gel buffer1.25ml10% SDS50ulddH2O1.42ml10% Ammonium persulfate*30ulTEMED*3 ul* Added after deaeration4% Stacking gel solution (2.5ml)30% Acrylamide solution332.5ul4 X stacking gel buffer625ul10% SDS25u
2、lddH2O1500ul10% Ammonium persulfate*15ulTEMED*2ul(常温保存)10% SDS (十二烷基硫酸钠)(4保存)10%AP (过硫酸镂)TEMED (N,NN,N四甲基乙二胺)4 X resolting gel buffer pH=8.80 175mlTris31.7639gdd H2O150mlHC1 (注:浓 hcd调pH至8.80ddH2O定容至175ml4 X stacking gel buffer pH=6.80 100mlTris(MW 121.1)6.0gdd H2O80mlHC1 (先用浓HCl,再用稀HCl调)pH6.80ddH2O至
3、 100ml30% Acrylamide solution 100mlAcrylamide (丙烯酰胺)30g甲叉双丙烯酰胺0.8gAdd ddH2O 37 溶解ddH2Oto 100ml(常温保存)电泳缓冲液(内槽)5 X Tank buffer 100mlTris 0.125M7.575gGlycine (甘氨酸)36.03gSDS2.5gddH2Oto 500ml使用前稀释10倍电泳缓冲液(外槽)使用过的内槽电泳缓冲液固定液取5()%甲醇454mL冰乙酸46ml混匀考马斯亮蓝R-250400mlR-2500.485g无水乙醇95mlddH2Oto 4(X)ml脱色液冰乙酸75mL甲醇50ml,加蒸饰水定容至l()(X)ml