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nvwangzzh

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女王

[求助] 急急急,投的是Phytomedicine,给了意见,有些看不懂,请高人指点 已有1人参与

In silico studies using molecular docking approaches are not stand-alone techniques and results have to be experimentally validated in vitro. Otherwise they remain hypothetical. Without functional assays (e.g. enzymatic inhibition) and binding studies (e.g. by surface plasmon resonance, isothermal calorimetry or microscale thermopheresis), there is no evidence, whether or not in silico results are correct. Unfortunately, the in vitro validation of the presented in silico data is missing in the present manuscript.   请问   functional assays (e.g. enzymatic inhibition) and binding studies (e.g. by surface plasmon resonance, isothermal calorimetry or microscale thermopheresis),是什么实验呢?


There should be in vitro kinase assays performed to examine the inhibitory activities of convallatoxin on JAKs, Src, and PI3K.  请问 vitro kinase assays是什么实验呢?
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zsw2366

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in vitro kinase assay 是指在体外条件下(适当的缓冲体系),采用1.重组的激酶(JAKs, Src, and PI3K.)与这些激酶适当的2.蛋白底物,加上放射性标记ATP[磷32],在无或者有抑制剂的条件下孵育一段时间之后,跑蛋白电泳。将跑完电泳的胶,与X光片共同孵育一段时间,几天到一周不等。如果你的蛋白底物发生磷酸化,则被P32标记,在该底物分子量的位置,将会有放射性自显影条带。如果你的抑制剂起作用的话,响应条带的强弱将降低。
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