Miskolci Veronika, Spiering Désirée, Cox Dianne, Hodgson Louis
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Ave, Bronx, NY, 10461, USA,
Methods Mol Biol. 2014;1172:173-84. doi: 10.1007/978-1-4939-0928-5_15.
Cytokine stimulations of leukocytes many times result in transient activation of the p21 Rho family of small GTPases. The role of these molecules during cell migration and chemotaxis is well established. The traditional approach to study the activation dynamics of these proteins involves affinity pull-downs that are often cumbersome and prone to errors. Here, we describe a reagent and a method of simple "mix-and-measure" approach useful for determining the activation status of endogenous Cdc42 GTPase from cell lysates.
细胞因子对白细胞的多次刺激常常导致小GTP酶的p21 Rho家族的瞬时激活。这些分子在细胞迁移和趋化作用中的作用已得到充分证实。研究这些蛋白质激活动力学的传统方法涉及亲和下拉，这种方法通常很繁琐且容易出错。在这里，我们描述了一种试剂和一种简单的“混合并测量”方法，可用于确定细胞裂解物中内源性Cdc42 GTP酶的激活状态。
