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文献和实验Rac and Rap GTPase Activation Assays
by the development of high-affinity probes for small GTPases. These probes are created by fusing a high-affinity GTPase-binding domain derived from a specific downstream effector protein to glutathione-S -transferase (GST). Such domains bind preferentially
(GST)或者是 GST fusion protein包含了Cdc42/Rac binding domain (CRIB) of PAK1 immobilized on glutathione–Sepharose 4B beads. The beads 用 lysis buffer清洗三次, bound GTP-Rac1 采用Rac1抗体进行免疫印渍分析 就我目前的了解来说,Rac和Cdc42都是可以采取这样的方式来检测其活性的.
Ubiquitination of Rac1 by Inhibitors of Apoptosis (IAPs)
that inhibitors of apoptosis (IAPs) function as direct E3 ubiquitin ligases for Rho GTPase Rac1 and target it for proteasomal degradation. Here, we describe in vitro and in vivo ubiquitination assays for detecting the conjugation
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