Cyclic AMP XP? Chemiluminescent Assay Kit

产品编号：CST-8019S 品牌：CST 原厂货号：8019S
产品分类：生化和细胞分析 > 其它细胞分析试剂盒 > 其它细胞分析试剂盒 > 其它
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包装：

运保温度:

-20° C

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其它组分：

ELISA Wash Buffer (20X)【包装：10 ml，，运保温度：-20° C】

Cell Lysis Buffer (10X) 【子货号：#9803，包装：15 ml，，运保温度：-20° C】

Sealing Tape【包装：2 sheets，，运保温度：-20° C】

Stable Peroxide Buffer【包装：3 ml，，运保温度：-20° C】

Luminol/Enhancer Solution【包装：3 ml，，运保温度：-20° C】

cAMP Ab Coated Microwells【包装：96 tests，，运保温度：-20° C】

cAMP-HRP Conjugate【包装：5.5 ml，，运保温度：-20° C】

cAMP Standard (2.4uM)【包装：0.5 ml，，运保温度：-20° C】

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。相对小的交叉反应在cGMP和cIMP中观察到，而cAMP的敏感性比此两者高10倍。在测试的各物质中没有观察到其它交叉反应。试剂盒对cAMP的敏感性，如图1所示，范围为0.2-12nM。特异性处理后的细胞cAMP水平的变化如图2（CHO细胞）所示。Cyclic AMP XP Chemiluminescent 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cAMP水平。此实验中，cAMP被发现在测试样本中与固定数量的HRP连接的cAMP竞争结合固定在96孔板上的抗cAMP XP兔单克隆抗体。接下来的洗涤移除了多余的样本cAMP和HRP连接的cAMP，然后加入Chemiluminescent（化学发光剂）以显色。由于此法的竞争性，使用相对发光单位（RLU）测量的发光强度与样本中cAMP的数量呈反比。使用cAMP标准溶液进行发光值测量，可计算出目标样本中cAMP的绝对数量。环磷酸腺苷3'-5'-单磷酸酶（cAMP）是一个重要的第二信使，在不同类型的细胞中涉及许多信号转导通路（1-3）。在哺乳动物细胞中，这一重要分子是由腺苷酸环化酶（AC）产生的。细胞外刺激如神经递质，激素，趋化因子，脂类介质和药物，可以通过结合大量跨膜G蛋白偶联受体调节AC活性以增加或减少cAMP的产生（4）。cAMP降解为AMP由磷酸酶催化，通过细胞内核苷酸浓度、磷酸化或结合钙/钙调蛋白及其它蛋白质来调节（5）。一组不同的分子，包括cAMP依赖的蛋白激酶PKA，环核苷酸门控的离子通道以及交换蛋白均可被cAMP（Epac）激活而介导下游cAMP信号通路（6,7）。cAMP调制多种生物学过程，包括代谢、分化、心肌细胞功能、神经信号转导、细胞黏附和免疫功能（5-7）。

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. Relatively minor cross-reactivity was observed with cGMP and cIMP, with 10 fold higher sensitivity for cAMP compared to either cGMP or 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 0.2 to 12 nM of cAMP. Changes in cellular cAMP levels following specific treatment with forskolin is shown in Figure 2 (CHO cells).

Description

The Cyclic AMP XP® Chemiluminescent Assay Kit is a competition enzyme-linked immunoassay used to determine cAMP levels in cells or tissues of interest. In this assay, cAMP found in the test sample competes with a fixed amount of HRP-linked cAMP for binding to an anti-cAMP XP® Rabbit mAb immobilized onto a 96-well plate. Following washing to remove excess sample cAMP and HRP-linked cAMP, chemiluminescent reagent is added for signal development. Because of the competitive nature of this assay, the magnitude of light emission, measured in relative light units (RLU), is inversely proportional to the quantity of sample cAMP. Measurement of light emmision using the cAMP Standard allows calculating the absolute amount of cAMP in a sample of interest.

Background

Cyclic adenosine 3’,5’-monophosphate (cAMP) is an important second messenger involved in many signal transduction pathways in different cell types of numerous species (1-3). In mammalian cells, this important molecule is produced by adenylyl cyclases (AC). Extracellular stimuli such as neurotransmitters, hormones, chemokines, lipid mediators, and drugs can modulate AC activity to increase or decrease cAMP production by binding to a large number of transmembrane G protein-coupled receptors (4). The degradation of cAMP to AMP is catalyzed by phosphodiesterases that are regulated by intracellular nucleotide concentrations, phosphorylation, or binding of Ca2+/calmodulin and other regulatory proteins (5). A set of diverse molecules, including cAMP-dependent protein kinase (PKA), cyclic nucleotide-gated ion channels, and exchange proteins that are activated by cAMP (EPAC), mediate downstream cAMP signaling (6,7). cAMP modulates various biological processes including metabolism, differentiation, cardiac cell functions, neuronal signaling, cell adhesion, and immune functions (5-7).

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.