PathScan? Total VEGFR-2 Sandwich ELISA Kit

• 产品编号：CST-7340C      品牌：CST       原厂货号：7340C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

CST公司的PathScan® Total VEGFR-2 Sandwich ELISA Kit主要用于检测内源性VEGFR-2的表达。如图1所示，用PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA Kit #7335能够检测到VEGF处理后HUVE*细胞中磷酸化VEGFR-2的含量显著升高，但用Sandwich ELISA Kit #7340检测到的总VEGFR-2（磷酸化的和未磷酸化的）的量不变。内部检测显示，本试剂盒能检测列举出的物种蛋白，但不排除用于其它物种同源蛋白检测的可能。*人脐静脉内皮细胞。CST公司的PathScan® Total VEGFR-2 Sandwich ELISA Kit利用固相双抗夹心酶联免疫检测法对内源性总VEGFR-2蛋白的表达进行检测。微孔中已经包被了VEGFR-2 鼠单抗 (7335-1D6*)，与细胞裂解液共孵育后，磷酸化的和未磷酸化的VEGFR-2均能被包被抗体捕获。多次洗板之后，加入VEGFR-2兔单抗 (7340-55B11*)对被捕获的VEGFR-2蛋白进行检测，然后加入HRP标记的抗兔抗体 (#7074*)对检测抗体进行识别，最后加入HRP的底物TMB进行显色反应。按照相对光单位，发射光强度与VEGFR-2蛋白的量成正比。*试剂盒中的抗体为试剂盒特异定制配方。血管内皮生长因子受体2 (VEGFR2, KDR, Flk-1)是内皮细胞上VEGF诱导信号时的一个主要受体。和配体结合之后，VEGFR2发生自磷酸化并被活化(1)。VEGFR2 主要的自磷酸化受体位于激酶插入区 (Tyr951/996)和酪氨酸激酶催化区(Tyr1054/1059) (2)。受体的激活会导致对包括Shc, Grb2, PI3K激酶, Nck 和蛋白酪氨酸磷酸化酶SHP-1 和 SHP-2接头蛋白的快速招募(3)。Tyr1212的磷酸化为Grb2提供了结合位点而且磷酸化的Tyr1175能结合PI3K激酶的p85 亚基以及PLCγ和 Shb (1,4,5)。VEGFR2信号传导对于实现VEGF刺激引起的增殖、趋化和发芽，以及体外培养的内皮细胞的增殖和体内血管的发生是必需的(6-8)。

1. Meyer, M. et al. (1999) EMBO J 18, 363-74.
2. Dougher-Vermazen, M. et al. (1994) Biochem Biophys Res Commun 205, 728-38.
3. Kroll, J. and Waltenberger, J. (1997) J Biol Chem 272, 32521-7.
4. Takahashi, T. et al. (2001) EMBO J 20, 2768-78.
5. Holmqvist, K. et al. (2004) J Biol Chem 279, 22267-75.
6. Karkkainen, M.J. and Petrova, T.V. (2000) Oncogene 19, 5598-605.
7. Rahimi, N. et al. (2000) J Biol Chem 275, 16986-92.
8. Claesson-Welsh, L. (2003) Biochem Soc Trans 31, 20-4.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Total VEGFR-2 Sandwich ELISA Kit detects endogenous levels of total VEGFR-2 protein. Using PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA Kit #7335, a significant induction of phospho-VEGFR-2 in HUVE* cells treated with VEGF can be detected. However, the level of total VEGFR-2 (phospho and non-phospho), detected by this Sandwich ELISA Kit #7340, remains unchanged (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. 
 *Human Umbilical Vein Endothelial Cells

Description

CST's PathScan® Total VEGFR-2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total VEGFR-2 protein. A VEGFR-2 Mouse mAb (7335-1D6*) has been coated onto the microwells. After incubation with cell lysates, Both nonphospho- and phospho-VEGFR-2 proteins are captured by the coated antibody. Following extensive washing, a VEGFR-2 Rabbit mAb (7340-55B11*) is added to detect the captured VEGFR-2 protein. HRP-linked anti-rabbit antibody (#7074*) is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total VEGFR-2 protein. 
 * Antibodies in this kit are custom formulations specific to the kit.

Background

Vascular endothelial growth factor receptor 2 (VEGFR2, KDR, Flk-1) is a major receptor for VEGF-induced signaling in endothelial cells. Upon ligand binding, VEGFR2 undergoes autophosphorylation and becomes activated (1). Major autophosphorylation sites of VEGFR2 are located in the kinase insert domain (Tyr951/996) and in the tyrosine kinase catalytic domain (Tyr1054/1059) (2). Activation of the receptor leads to rapid recruitment of adaptor proteins, including Shc, Grb2, PI-3 kinase, Nck and the protein tyrosine phosphatases SHP-1 and SHP-2 (3). The phosphorylation of Tyr1212 provides a docking site for Grb2 binding and phospho-Tyr1175 binds with the p85 subunit of PI-3 kinase and PLCγ, as well as Shb (1,4,5). Signaling from VEGFR2 is necessary for the execution of VEGF-stimulated proliferation, chemotaxis and sprouting, as well as survival of cultured endothelial cells in vitro and angiogenesis in vivo (6-8).

1. Meyer, M. et al. (1999) EMBO J 18, 363-74.
2. Dougher-Vermazen, M. et al. (1994) Biochem Biophys Res Commun 205, 728-38.
3. Kroll, J. and Waltenberger, J. (1997) J Biol Chem 272, 32521-7.
4. Takahashi, T. et al. (2001) EMBO J 20, 2768-78.
5. Holmqvist, K. et al. (2004) J Biol Chem 279, 22267-75.
6. Karkkainen, M.J. and Petrova, T.V. (2000) Oncogene 19, 5598-605.
7. Rahimi, N. et al. (2000) J Biol Chem 275, 16986-92.
8. Claesson-Welsh, L. (2003) Biochem Soc Trans 31, 20-4.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately)