PathScan? PDGFR Activity Assay: Phospho-PDGFR, Phospho-SHP2, Phospho-Akt, and Phospho-p44/42 MAPK (E

• 产品编号：CST-5304S      品牌：CST       原厂货号：5304S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

cocktail中的每个磷酸化抗体都只能识别内源性靶蛋白的磷酸化形式。eIF4E antibody能检测内源性靶蛋白水平，并且不依赖于磷酸化，同时该抗体还能用于蛋白负载对照。该抗体通过使用合成肽段免疫动物而获得，并经蛋白A和肽亲和层析纯化。PathScan® Multiplex Western Detection Cocktail为分析一张膜上多种蛋白的活性状态提供了独特的方法，不用去除免疫印迹膜上的封闭液和抗体，然后再次重新标记。该方法为使用者节约了时间，且调高了准确度，减少了试剂的浪费。PDGFR Tyrosine Kinase Activity Assay让使用者能即时检测DGF受体, SHP2, Akt和p44/42 MAPK蛋白应答PDGF后的磷酸化。该试剂盒也包括用于控制蛋白负载的eIF4E Antibody。血小板衍生生长因子(PDGF)是一种二聚体分子，它以相关多肽链的同源二聚体或异源二聚体形式存在(A和B)，目前已经鉴定出两种PDGF受体。PDGFalpha-受体结合高亲和性地结合三种异构体，然而beta-受体只能高亲和性地结合PDGF-BB，低亲和性地结合PDGF-AB，并不结合PDGF-AA (1)。PDGF通过结合这些两种相关蛋白酪氨酸激酶受体而发挥它对细胞的刺激效应。配体的结合诱导受体的二聚化和自磷酸化，使胞质内含有SH2结构域的信号转导分子结合、活化。因此，许多不同的信号转导通路被开启，导致细胞生长、肌动蛋白重组、迁移和分化(2-4)。在临床研究中，人们发现PDGF的表达出现在许多不同的实质固态瘤中，从恶性胶质瘤到前列腺癌。在这些不同的肿瘤中，PDGF信号转导的生理作用各不相同，它对癌细胞生长的自分泌刺激作用不同于涉及毗邻间质甚至血管生成的更多敏感旁分泌相互作用。靶点PDGF信号转导已经成为肿瘤治疗的一个有效方法(5)。

1. Westermark, B. et al. (1990) Ciba Found. Symp. 150, 6-22.
2. Heldin, C.H. (1997) FEBS Lett. 410, 17-21.
3. Bornfeldt, K. E. et al. (1995) Ann. N. Y. Acad. Sci. 766, 416-430.
4. Renhowe, P.A. (2002) Curr. Opin. Drug Discov. Devel. 5, 214-224.
5. George, D. (2001) Semin. Oncol. 28, 27-33.

原厂资料：

Specificity / Sensitivity

Each phospho-antibody in this cocktail recognizes endogenous levels of only the phosphorylated form of its specific target. The eIF4E antibody detects endogenous levels of its target independent of phosphorylation and is provided to control for protein loading.

Source / Purification

Antibodies are produced by immunizing animals with synthetic peptides, and are purified by combinations of Protein A and peptide affinity chromatography.

Description

The PathScan® Multiplex Western Detection Cocktail offers a unique method to assay the activation of multiple proteins on one membrane without stripping and reprobing. This method saves the user valuable time while increasing accuracy and minimizing reagent waste. The PDGFR Tyrosine Kinase Activity Assay allows the user to simultaneously detect the phosphorylation of PDGF receptor, SHP2, Akt and p44/42 MAPK proteins in response to PDGF. The kit also includes an eIF4E antibody to control protein loading.

Background

Platelet-derived growth factor (PDGF) is a dimeric molecule that exists as homodimers or heterodimers of related polypeptide chains (A and B). Two types of PDGF receptors have been identified. The PDGF alpha-receptor binds all three isoforms with high affinity, whereas the beta-receptor binds only PDGF-BB with high affinity, PDGF-AB with low affinity and does not appear to bind PDGF-AA (1). PDGF exerts its stimulatory effects on cells by binding to these two related protein tyrosine kinase receptors. Ligand binding induces receptor dimerization and autophosphorylation, allowing binding and activation of cytoplasmic SH2-domain-containing signal transduction molecules. Thereby, a number of different signaling pathways are initiated, leading to cell growth, actin reorganization, migration and differentiation (2-4). In clinical studies, PDGF expression has been shown in a number of different solid tumors, from glioblastomas to prostate carcinomas. In these various tumor types, the biologic role of PDGF signaling can vary from autocrine stimulation of cancer cell growth to more subtle paracrine interactions involving adjacent stroma and even angiogenesis. Targeting PDGF signaling becomes an effective way for tumor treatment (5).

1. Westermark, B. et al. (1990) Ciba Found. Symp. 150, 6-22.
2. Heldin, C.H. (1997) FEBS Lett. 410, 17-21.
3. Bornfeldt, K. E. et al. (1995) Ann. N. Y. Acad. Sci. 766, 416-430.
4. Renhowe, P.A. (2002) Curr. Opin. Drug Discov. Devel. 5, 214-224.
5. George, D. (2001) Semin. Oncol. 28, 27-33.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not
aliquot the antibody