Active Arf1 Detection Kit

• 产品编号：CST-8816S      品牌：Cell Signaling Technology       原厂货号：8816S
• 产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞结构分析试剂盒
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描述：

Active Arf1 Detection Kit能够检测内源性的结合GTP(活化) 的Arf1水平，如图1所示。经过内部实验验证，该试剂盒能够检测指定物种的蛋白，但可能也可以检测其他物种的同源蛋白。Active Arf1 Detection Kit提供了检测细胞中Arf1 GTPase活性所必需的所有试剂。GST-GGA3-PBD融合蛋白用于结合活化形式的GTP结合的Arf1，该融合蛋白随后可被谷胱甘肽树脂免疫沉淀。之后通过western blot方法检测Arf1活化水平，使用的抗体是Arf1 Rabbit mAb兔单抗。Ras小GTP结合蛋白 (G 蛋白)超家族是由一大类蛋白（超过150个成员）组成，这些蛋白可根据他们的序列和功能相似性分成至少5个家族：Ras, Rho, Rab, Arf以及 Ran (1-3)。这些小G蛋白同时具有GDP/GTP结合活性和GTPase活性，在许多细胞和发育事件如细胞周期进程、细胞存活、肌动蛋白细胞骨架组装、细胞极性和运动以及囊泡和核转运中可作为双向开关（1）。上游信号可刺激GDP从GDP-bound形式（非活化）解离，从而导致GTP结合并形成GTP-boung形式（活化）。活化的G蛋白随后在其下游效应因子-结合区域发生一个构象改变，从而与下游效应因子结合并进行调控。这种激活作用可被其内在的GTPase活性关闭（GTPase可以水解GTP生成GDP）并释放下游效应因子。Ras超家族蛋白的这些固有的鸟苷酸交换因子以及GTP水解活性同样也可被鸟苷酸交换因子 (GEFs) 和GTPase激活蛋白(GAPs)调控，GEF可以促进活化的GTP-bound形式的形成，GAP可使GTPase返回至GDP-bound的非活化形式（4）。ADP-ribosylation factor (Arf)蛋白是一个属于Ras GTPase超家族的低分子量GTP结合蛋白。Arf1 蛋白可根据其氨基酸序列和结构的相似性被分成三种不同的种类。Arf1是I类Arf蛋白，参与囊泡从Glogi的出芽（1）。

1. Takai, Y. et al. (2001) Physiol Rev 81, 153-208.
2. Colicelli, J. (2004) Sci STKE 2004, RE13.
3. Wennerberg, K. et al. (2005) J Cell Sci 118, 843-6.
4. Vigil, D. et al. (2010) Nat Rev Cancer 10, 842-57.

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Specificity / Sensitivity

Active Arf1 Detection Kit detects endogenous levels of GTP-bound (active) Arf1 as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The Active Arf1 Detection Kit provides all reagents necessary for measuring activation of Arf1 GTPase in the cell. GST-GGA3-PBD fusion protein is used to bind the activated form of GTP-bound Arf1, which can then be immunoprecipitated with glutathione resin.Arf1 activation levels are then determined by western blot using an Arf1 Rabbit mAb.

Background

The Ras superfamily of small GTP-binding proteins (G proteins) comprise a large class of proteins (over 150 members) that can be classified into at least five families based on their sequence and functional similarities: Ras, Rho, Rab, Arf, and Ran (1-3). These small G proteins have both GDP/GTP-binding and GTPase activities and function as binary switches in diverse cellular and developmental events that include cell cycle progression, cell survival, actin cytoskeletal organization, cell polarity and movement, and vesicular and nuclear transport (1). An upstream signal stimulates the dissociation of GDP from the GDP-bound form (inactive), which leads to the binding of GTP and formation of the GTP-bound form (active). The activated G protein then goes through a conformational change in its downstream effector-binding region, leading to the binding and regulation of downstream effectors. This activation can be switched off by the intrinsic GTPase activity, which hydrolyzes GTP to GDP and releases the downstream effectors. These intrinsic guanine nucleotide exchange and GTP hydrolysis activities of Ras superfamily proteins are also regulated by guanine nucleotide exchange factors (GEFs) that promote formation of the active GTP-bound form and GTPase activating proteins (GAPs) that return the GTPase to its GDP-bound inactive form (4).ADP-ribosylation factor (Arf) proteins are low molecular weight GTP binding proteins that belong to the Ras GTPase superfamily. Arf proteins are grouped into three distinct classes based on amino acid sequence and structural similarity. Arf1 is a class I Arf protein and is involved in the budding of vesicles from the Golgi (1).

1. Takai, Y. et al. (2001) Physiol Rev 81, 153-208.
2. Colicelli, J. (2004) Sci STKE 2004, RE13.
3. Wennerberg, K. et al. (2005) J Cell Sci 118, 843-6.
4. Vigil, D. et al. (2010) Nat Rev Cancer 10, 842-57.