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Author:
Date:
2014-03-05
[Abstract] Immunoprecipitation (IP)- Kinase assays are an invaluable tool to assess the activation status of intracellular signaling cascades within a specific cellular state and also to confirm the enzymatic activity of a specific kinase towards a putative substrate of interest. Intracellular signal transduction cascades play an important role in modulating the localization of transcription factors and thus impact the cellular transcriptome. This in turn regulates key cell fate decisions including cell survival, apoptosis, proliferation, and differentiation. Here we describe an in vitro non-radioactive method to assess kinase activity towards a specific substrate. In this protocol we outline the method for Akt, however the basic protocol may be applied to any kinase and putative substrate ...
[摘要] 免疫沉淀（IP） - 激酶测定是评估特定细胞状态内的细胞内信号级联的活化状态以及确认特异性激酶朝向目的基因的酶活性的有价值的工具。 细胞内信号转导级联在调节转录因子的定位中发挥重要作用，从而影响细胞转录组。 这反过来调节关键细胞命运决定，包括细胞存活，凋亡，增殖和分化。 在这里，我们描述了一种体外非放射性方法来评估对特定底物的激酶活性。 在这个协议中，我们概述了Akt的方法，但是基本协议可以应用于任何激酶和感兴趣的推定底物。
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