PathScan? RTK Signaling Antibody Array Kit (Fluorescent Readout)

• 产品编号：CST-7949S      品牌：CST       原厂货号：7949S
• 产品分类：ELISA及其它免疫分析 > 其它免疫检测 > 抗体芯片（Antibody Array)
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描述：

Cell Signaling Technology公司的PathScan® RTK Signaling Antibody Array Kit 仅在酪氨酸或其它特殊位点磷酸化（见实验靶点图）时用于检测RTKs和信号结点。除了FLT3抗体和磷酸化的EphB3有些交叉反应外，未见靶点间有明显交叉反应。此外， Stat1 (Tyr701) 和Stat3 (Tyr705)在其蛋白的其它酪氨酸位点磷酸化时也能被检测到。本试剂盒在细胞裂解液的总蛋白含量在0.2 和1 mg/ml 时效果最好(见图4)。所用的捕获抗体对人的靶点有效。虽然本试剂盒未在小鼠裂解液中进行过测试，但很多捕获抗体可能与小鼠的体系发生交叉反应。PathScan® RTK Signaling Antibody Array Kit (荧光法检测) 是一个基于双抗夹心免疫检测原理的玻璃板抗体芯片。本芯片能在酪氨酸位点或其它位点磷酸化时同时检测28种酪氨酸激酶受体和11种重要的信号结点。靶点特异的捕获抗体，生物素化的蛋白（阳性对照）和非特异IgG (阴性对照) 已经重复点在了硝酸纤维包被的玻璃板上。每个试剂盒含8张芯片，能检测16个样品。细胞裂解液和芯片共孵育后，再和生物素标记的检测抗体组合反应。然后用链霉亲和素标记的 DyLight 680® 使检测抗体可视化。芯片的荧光成像能被数码成像系统捕获并能用芯片分析软件对点密度进行定量。受体酪氨酸激酶(RTKs) 是一个主要通过酪氨酸磷酸化传递信号的细胞表面受体家族(1)。 RTKs 能激发包括PI3K/Akt, MAPK 和 Jak/Stat通路在内的许多下游信号。这些通路控制着分裂、生长、代谢、分化、迁移和存活等许多基本的细胞活动。RTK信号失调在大量的肿瘤中被发现(2)，使得RTKs 成为药物研发的热门靶点。

1. Schlessinger, J. (2000) Cell 103, 211-25.
2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
3. Looyenga, B.D. et al. (2012) PLoS One 7, e30820.

原厂资料：

Specificity / Sensitivity

Cell Signaling Technology's PathScan® RTK Signaling Antibody Array Kit detects the indicated RTKs and signaling nodes only when phosphorylated at tyrosine or specified residues (see Array Target Map). No significant crossreactivity has been observed between targets, with the exception of some crossreactivity of the FLT3 antibody with phosphorylated EphB3. In addition, Stat1 (Tyr701) and Stat3 (Tyr705) may be detected when phosphorylated at other tyrosine sites within the proteins. This kit is optimized for cell lysates diluted to a total protein concentration between 0.2 and 1 mg/ml (see Figure 4). All capture antibodies have been validated for human targets. Although this kit has not been tested with mouse lysates, it is expected than many capture antibodies will crossreact in murine systems.

Description

The PathScan® RTK Signaling Antibody Array Kit (Fluorescent Readout) is a slide-based antibody array product founded upon the sandwich immunoassay principle. The array kit allows for the simultaneous detection of 28 receptor tyrosine kinases and 11 important signaling nodes when phosphorylated at tyrosine or other residues. Target-specific capture antibodies, biotinylated protein (positive control) and nonspecific IgG (negative control) have been spotted in duplicate onto nitrocellulose-coated glass slides. Each kit contains two 8-pad slides, allowing the user to test up to 16 samples. Cell lysate is incubated on the slide followed by a biotinylated detection antibody cocktail. Streptavidin-conjugated DyLight 680® is then used to visualize the bound detection antibody. A fluorescent image of the slide can then be captured with a digital imaging system and spot intensities quantified using array analysis software.

Background

Receptor Tyrosine Kinases (RTKs) are a family of cell surface receptors that signal primarily through tyrosine phosphorylation events (1). RTKs trigger a wide range of downstream signaling cascades, including the PI3K/Akt, MAPK and Jak/Stat pathways. These pathways control basic cellular functions such as division, growth, metabolism, differentiation, migration and survival. Dysregulation of RTK signaling has been implicated in a large number of cancers (2), making RTKs popular targets for pharmaceutical intervention.

1. Schlessinger, J. (2000) Cell 103, 211-25.
2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
3. Looyenga, B.D. et al. (2012) PLoS One 7, e30820.

注意事项：

*Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).