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RhoA活性检测试剂盒
RhoA活性检测试剂盒图片
货号: 80601
品牌: NewEast Biosciences
参考价格: 6800元/盒
交货期: 现货


包装与价格:
订货号 纯度规格 包装 价格(元)
80601 30 Assays 6800
半岛bd体育手机客户端 名称
RhoA Pull-Down Assay Kit
半岛bd体育手机客户端 介绍
品牌:美国 NewEast Biosciences
货号:80601

规格:30 Assays
半岛bd体育手机客户端 全名:RhoA Pull-Down Assay Kit
应用:Pull-Down



监测:Active RhoA-GTP levels


RhoA Pull-Down Activation Assay Kit

Cat. # 80601

Introduction

A. Background
Small GTPases are a super-family of cellular signaling regulators. RhoA belongs to the Rho sub-family of GTPases that regulate cell motility, cell division, and gene transcription. GTP binding increases the activity of RhoA, and the hydrolysis of GTP to GDP renders it inactive.
Currently the activation of RhoA proteins is assayed with the binding of GTP-bound RhoA to the p21-binding domain (PBD) of p21-activated protein kinase (PAK). This method is based on the observation that the active, GTP-bound RhoA could bind to the PBD of PAK. However, the reproducibility of this method is poor. This is partially due to the relatively quick hydrolysis of GTP to GDP during the assay procedure, and the low binding affinity of PBD to RhoA-GTP.
The RhoA Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes RhoA-GTP, but not RhoA-GDP. Given the high affinity of monoclonal antibodies to their antigens, the activation assay could be performed in a much shorter time. This assay provides the reliable results with consistent reproducibility.
The anti-RhoA-GTP monoclonal antibody can also be used to monitor the activation of RhoA in cells and in tissues by immunohistochemistry.
B. Assay Principle
The RhoA Activation Assay Kit uses configuration-specific anti-RhoA-GTP Mouse monoclonal antibody to measure RhoA-GTP levels in cell extracts or in vitro GTPγS loading RhoA activation assays. Anti-RhoA-GTP mouse monoclonal antibody is first incubated with cell lysates containing RhoA-GTP. Next, the GTP-bound RhoA is pulled down by protein A/G agarose. Finally, the precipitated RhoA-GTP is detected through immunoblot analysis using Anti-RhoA Rabbit Polyclonal Antibody.
C. Kit Components
1. Anti-RhoA-GTP Mouse Monoclonal Antibody (Cat. # 26904): One vial – 35 μL (1 mg/ml) in PBS, pH 7.4, containing 50% glycerol. This antibody specifically recognizes RhoA-GTP from all vertebrates.
2. Protein A/G Agarose (Cat. # 30301): One vial – 600 μL of 50% slurry.
3. 5X Assay/Lysis Buffer (Cat. # 30302): One bottle – 30 mL of 250 mM Tris-HCl, pH 8, 750 mM NaCl, 50 mM MgCl2, 5 mM EDTA, 5% Triton X-100.
4. Anti-RhoA Rabbit Polyclonal Antibody (Cat. # 21017): One vial – 50 μL (1 mg/mL) in PBS, pH 7.4, contained 50% glycerol.
5. 100X GTPγS (Cat. # 30303): One vial – 50 μl at 10 mM, use 5 μL of GTPγS for GTP-labeling of 0.5 mL of cell lysate.
6. 100X GDP (Cat. # 30304): One vial – 50 μl at 100 mM, use 5 μL of GDP for GDP-labeling of 0.5 mL of cell lysate.
7. HRP-Goat Anti-Rabbit IgG (Cat. #29002): 50 μL (0.4 mg/mL) in PBS, pH 7.4, contained 50% glycerol.


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