SDS-PAGE电泳初级指南.docx

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% SDS25ulddH2O1500ul10% 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 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