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HX91926

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[求助] 请问镍亲和纯化后的蛋白液总是负值怎么回事

上一次做纯化,流完40mM的咪唑,用100mM的开始洗,结果没洗几管就开始变负值了,而且还挺大的,都是-0.030,-0.040这么大
这次做,我洗40的,结果洗到30多管,又流了十几分钟还是0.020这么大,我就换浓度了,由于上次100的是负值,所以这次直接用250的洗,结果还是没几管就负值了,而且负值也都挺大(每次换浓度,我有换对照的),请问怎么回事
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