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Merck
CN

17-441

Sigma-Aldrich

Rac1/Cdc42活化检测试剂盒

The Rac1/Cdc42 Activation Assay provides an effective method for detecting Rac & Cdc42 activity in cell lysates.

别名:

Rac1激活检测

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关于此项目

UNSPSC代码:
12161503
eCl@ss:
32161000
NACRES:
NA.84
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质量水平

种属反应性

human, mouse, rat

制造商/商品名称

Upstate®

技术

activity assay: suitable
affinity binding assay: suitable (G-protein)

UniProt登记号

运输

wet ice

基因信息

human ... RAC1(5879)

一般描述

Rac1/Cdc42活化测定法(目录编号17-441)提供了检测细胞裂解物中Rac和Cdc42活性的有效方法。 该测定法使用Rac/Cdc42的下游效应子p21激活的蛋白激酶(PAK1)从样品中分离Rac/Cdc42的活性GTP结合形式。 PAK1的p21结合域(PBD)表示为GST融合蛋白,并与琼脂糖珠偶联。 沉淀后,进行免疫印迹,并使用特异性单克隆抗体检测活化的Rac/Cdc42,然后使用HRP偶联的二抗和ECL试剂检测。

生化/生理作用

物种交叉反应性: 人和小鼠。预测与所有哺乳动物物种发生交叉反应。
抗-Rac1,克隆23A8:物种交叉反应性:人类,小鼠和大鼠。其他物种交叉反应性未知。
抗-cdc42 物种交叉反应性: 人,小鼠,大鼠和狗。 其他物种交叉反应性未知。

制备说明

自发货之日起在-20°C下保存1年

分析说明

通过从已加载GTPγS的3T3/A31和HeLa细胞裂解物中沉淀Rac1-GTP和cdc42-GTP进行常规评估(裂解物在体外用GTPγS加载cdc42-GTP)。 使用抗Rac1(1 μg/mL)通过免疫印迹分析检测沉淀的Rac1-GTP和cdc42-GTP。

其他说明

Rac/cdc42测定试剂(PAK-1 PBD,琼脂糖),目录号14-325:一个小瓶,含有300 μg PAK-1 PBD与150 μL谷胱甘肽琼脂糖微珠结合,以50%的20mM PBS(pH值7.4)浆液形式提供,含有50%的甘油,最终体积为300 μL。

抗-Rac1,克隆23A8,目录号05-389:一个小瓶,含有250 μg蛋白G纯化的小鼠IgG2b,溶于250 μL储存缓冲液(0.1 M Tris-甘氨酸,pH 7.4,0.15 M NaCl,含有0.05%叠氮化钠)中。

抗cdc42,(小鼠单克隆IgG1),目录号05-542:一小瓶,含有50 μg纯化小鼠IgG1,溶于200 μL 50%储存缓冲液(20 mM磷酸钠、pH 7.5、150 mM NaCl、1.5 mM叠氮化钠,含1 mg/mL BSA)和50%甘油中。

Mg2+裂解/清洗缓冲液,5X,目录号20-168:两个小瓶,每瓶包含18 mL的5X MLB:125 mM HEPES、pH 7.5、750 mM NaCl、5%Igepal CA-630、50 mM MgCl2、5 mM EDTA和10%甘油。

100X GTPγS,10 mM,目录号20-176:一个小瓶,包含50 μL的10 mM GTPγS,100X储备液,溶于50 mM Tris-HCl,pH 7.8,GTP的不可水解类似物中。 足以标记5 mL细胞裂解物。

100X GDP,100mM,目录号20-177:一个小瓶,含有50 μL 100 mM GDP、100X储备液,溶于50 mM Tris-HCl、pH值7.8中。 GDP(鸟苷5′-二磷酸)用于体外标记非活性形式的G蛋白。 足以标记5ml细胞裂解物。

法律信息

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

免责声明

除非我们的产品目录或产品附带的其他公司文档另有说明,否则我们的产品仅供研究使用,不得用于任何其他目的,包括但不限于未经授权的商业用途、体外诊断用途、离体或体内治疗用途或任何类型的消费或应用于人类或动物。

象形图

CorrosionEnvironment

警示用语:

Danger

危险声明

危险分类

Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1

储存分类代码

10 - Combustible liquids


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