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NuPAGETM NovexTM 4-12% Bis-Tris Protein Gels

NuPAGE TM Novex TM 4-12%Bis-Tris蛋白凝胶

Company: Thermo Fisher Scientific
Catalog#: NP0323PK2
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In vitro Microtubule Binding Assay and Dissociation Constant Estimation
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2016-03-20
[Abstract]  Microtubules (MTs) support an astonishing set of versatile cellular functions ranging from cell division, vesicle transport, and cell and tissue morphogenesis in various organisms. This versatility is in large mediated by MT-associated proteins (MAPs). The neuronal MAP Tau, for example, is stabilizing MTs in axons of the vertebrate nervous system and thus provides the basis for enduring axonal transport and the long life span of neurons (Mandelkow et al., 1994). Tau has been shown to bind to MTs directly in vitro and also to promote their nucleation from α-/β-tubulin subunits (Goode et al., 1994). Recently, we identified a plant-specific protein family called “companion of cellulose synthase” (CC), which was shown to bind MTs and enhance dynamics of the cortical ... [摘要]  微管(MT)支持惊人的一套多功能细胞功能,从细胞分裂,囊泡运输,以及细胞和组织形态发生在各种生物体。这种多功能性大量由MT相关蛋白(MAP)介导。例如,神经元MAP Tau是稳定脊椎动物神经系统轴突中的MT,因此为持久的轴突运输和神经元的长寿命提供了基础(Mandelkow等人,1994)。 Tau已经显示在体外直接结合MT,并且促进它们从α-/β-微管蛋白亚基的成核(Goode等人,1994)。最近,我们鉴定了称为"纤维素合酶的伴侣"(CC)的植物特异性蛋白质家族,其被证明结合MT并在盐胁迫下增强植物细胞中皮质MT阵列的动力学(Endler等,2015)。因此,CC被假定为帮助植物细胞应对胁迫条件,从而在不利生长条件下维持生物量生产。在这里,我们提供详细的实验信息在体外 MT结合测定,其允许评估感兴趣的蛋白质是否绑定到MT。该测定法在自旋向下方法中利用高分子量的MT,并且使得能够测定解离常数K d sub,其是蛋白质与MT的结合强度的量度。

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