Cyclic GMP XP? Assay Kit

• 产品编号：CST-4360S      品牌：CST       原厂货号：4360S
• 产品分类：ELISA及其它免疫分析 > 其它免疫检测 > 抗体芯片（Antibody Array)
• 应用分类：

其它组分：

描述：

此试剂盒的免疫活性经由以下物质测试：ADP, AMP, ATP, cAMP, cGMP, cIMP, cTMP, CTP, GDP, GMP以及GTP。在cIMP中观察到相对较小的交叉反应，但cGMP的敏感性是其100倍。在测试的各物质中没有观察到其它交叉反应。试剂盒对cGMP的敏感性，如图1所示，范围为2-200nM。 特异性处理后的细胞cGMP水平的变化如图2和图3（low passage RFL-6细胞）所示。Cyclic GMP XP Assay Kit是一个竞争性酶联免疫试验，用以分析目标细胞和组织的cGMP水平。此实验中，cGMP被发现在测试样本中与固定数量的HRP连接的cGMP竞争结合固定在96孔板上的抗cGMP XP兔单克隆抗体。接下来的洗涤移除了多余的样本cGMP和HRP连接的cGMP，然后加入HRP底物TMB以显色。由于此法的竞争性，显色值的强度与样本中cGMP的数量呈反比。使用cGMP标准溶液进行吸光值测量，可计算出目标样本中cGMP的绝对数量。注意：12个8孔板模组是设计用来进行分开的8次试验。环鸟苷酸3'-5'-单磷酸酶（cGMP）是一个重要的多功能第二信使，在几乎所有物种不同类型的细胞中涉及许多信号转导通路（1）。细胞内cGMP由鸟苷酸环化酶（GC）催化GTP形成，由磷酸二酯酶（PDE）水解降解（1,2）。GC的两个独特的家族已经确定：可溶性鸟苷酸环化酶（sGC），对一氧化氮反应并和细胞膜结合；颗粒鸟苷酸环化酶（pGC），对不同的细胞外激动剂反应，包括肽类激素，细菌毒素和自由基（2,3）。磷酸二酯酶由11个亚型形成一个超家族，对cAMP和cGMP的特异性各异（4）。cGMP通过激活cGMP依赖的激酶调节细胞生理功能，调节cGMP依赖的离子通道或转运体，通过PDE进行水解降解（1,4）。介于其具有多个效应分子，cGMP在调节多种生理和病理过程中扮演重要角色，例如血管平滑肌运动、肠液和电解质平衡以及视网膜光电转换（1,5）。

原厂资料：

Specificity / Sensitivity

The immunoreactivity of this kit was tested against the following: ADP, AMP, ATP, cAMP, cGMP, cIMP, cTMP, CTP, GDP, GMP and GTP. Minor cross-reactivity was observed with cIMP, with over 100 fold higher sensitivity for cGMP compared to cIMP. No cross-reactivity was observed with any of the other factors tested. Kit sensitivity, as shown in Figure 1, demonstrates a dynamic range of 2 to 200 nM of cGMP. Changes in cellular cGMP levels following specific treatments are shown in Figure 2 and Figure 3 (low passage RFL-6 cells).

Description

The Cyclic GMP XP® Assay Kit is a competition enzyme-linked immunoassay used to determine cGMP levels in cells or tissues of interest. In this assay, cGMP found in test sample competes with a fixed amount of HRP-linked cGMP for binding to an anti-cGMP XP® Rabbit mAb immobilized onto a 96-well plate. Following washing to remove excess sample cGMP and HRP-linked cGMP, HRP substrate TMB is added to develop color. Because of the competitive nature of this assay, the magnitude of the absorbance for this developed color is inversely proportional to the quantity of sample cGMP. Measurement of absorbance using the cGMP Standard allows calculating the absolute amount of cGMP in a sample of interest. 
 Note: 12 8-well modules -Each module is designed to break apart for 8 tests.

Background

Cyclic guanosine 3’,5’-monophosphate (cGMP) is a critical and multifunctional second messenger molecule involved in many signal transduction pathways in different cell types of almost all species (1). Intracellular cGMP is generated from GTP by guanylyl cyclase (GC) and degraded through phosphodiesterase (PDE) hydrolysis (1,2). Two distinctive families of GC have been identified: soluble guanylyl cyclases (sGC) that are nitric oxide-responsive and cell membrane-bound; and particulate guanylyl cyclases (pGC) that respond to diverse extracellular agonists including peptide hormones, bacterial toxins, and free radicals (2,3). Phosphodiesterases form a superfamily of 11 isoforms with different specificity to both cyclic adenosine 3’,5’-monophosphate (cAMP) and cGMP (4). Cyclic GMP regulates cellular physiology by activating cGMP-dependent kinase, modulating cGMP-dependent ion channels or transporters, and altering its own hydrolytic degradation by PDE (1,4). Because of the diversity of its effectors, cGMP plays an important role in regulating various pathological and physiological processes, such as vascular smooth muscle motility, intestinal fluid and electrolyte homeostasis, and retinal phototransduction (1,5).

注意事项：

Note: This kit contains mixed storage components. Please store this entire kit at -20° C for long term storage. Upon first
use, please allow components to thaw and then store each component as indicated on individual component labels.