Question Detail
When performing animal liver protein purification, can PMSF and cocktail be added together?
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Tags：#Phenylmethylsulfonyl fluoride #Protease inhibitor cocktail #Protein purification #Rat liver #Additive compatibility
All Answers4
A
AmyMedia7397
2026-09-19
杂带多不一定全因蛋白酶。先做对照：一组只加PMSF，一组加PMSF+cocktail，看杂带变化。若联合后杂带明显减少，说明存在多类蛋白酶，可常规联用；若差异不大，优先优化His纯化洗涤条件、咪唑浓度、盐浓度和核酸酶处理。
B
BioPro8665
2026-09-11
可以一起加，但先看cocktail说明书成分。有些cocktail已含AEBSF或丝氨酸蛋白酶抑制剂，再补PMSF要注意终浓度不要叠加过量。一般PMSF终浓度1mM，cocktail按1×。两者在裂解液体积中稀释后加入，一般不会产生沉淀；cocktail含DMSO时可能轻微浑浊，混匀后离心不影响后续使用。
谢
谢晨文
2026-09-10
不要将100mM PMSF母液与浓缩cocktail原液直接混合，可能因溶剂体系不同出现析出。正确做法是在冷的裂解液里先加一种混匀，再加另一种。PMSF和cocktail联合没有明显冲突，但PMSF对水不稳定，操作尽量冰上并在0.5-1小时内完成初步裂解。
C
Cell959
2026-07-02
顺序建议临用前先加PMSF到裂解液，混匀后再加cocktail。500mg肝脏按1:10加约5ml裂解液，100mM PMSF母液加50μl，cocktail按说明书加至1×。PMSF在水相半衰期短，裂解液放置超过30分钟可在纯化前少量补加，cocktail一般不需要频繁补。
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