| 货号 | 7340C |
| 描述 | CSTs 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. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs 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 |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | 4C |
| 参考文献 | Meyer, M. et al. (1999) EMBO J 18, 363-74. Dougher-Vermazen, M. et al. (1994) Biochem Biophys Res Commun 205, 728-38. Kroll, J. and Waltenberger, J. (1997) J Biol Chem 272, 32521-7. Takahashi, T. et al. (2001) EMBO J 20, 2768-78. Holmqvist, K. et al. (2004) J Biol Chem 279, 22267-75. Karkkainen, M.J. and Petrova, T.V. (2000) Oncogene 19, 5598-605. Rahimi, N. et al. (2000) J Biol Chem 275, 16986-92. Claesson-Welsh, L. (2003) Biochem Soc Trans 31, 20-4. |
Figure 1: Treatment of HUVE cells with VEGF stimulates phosphorylation of VEGFR-2 at Tyr1175, detected by PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA kit, #7335, but does not affect the level of total VEGFR-2 protein detected by PathScan® Total VEGFR-2 Sandwich ELISA kit, #7340. OD450 readings are shown in the top figure, while the corresponding Western blot using Phospho-VEGFR-2 (Tyr1175) Rabbit Monoclonal Ab #2478 (right panel) or VEGFR-2 Rabbit Monoclonal Antibody (7340-55B11) (left panel), is shown in the bottom figure.图1：用PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA kit, #7335试剂盒能够在VEGF刺激后的HUVE细胞中检测到VEGFR-2上Tyr1175位点的磷酸化，但用PathScan® Total VEGFR-2 Sandwich ELISA kit, #7340试剂盒检测总的VEGFR-2的量不变。上图显示OD450nm下的读值，下图显示用Phospho-VEGFR-2 (Tyr1175) Rabbit Monoclonal Ab #2478（右道）或VEGFR-2 Rabbit Monoclonal Antibody (7340-55B11)（左道）进行免疫印迹检测的结果。 | |
Figure 2: Linear relationship between protein concentration of lysates from VEGF-treated HUVE cells and kit assay optical density readings. HUVE cells (85% confluence) were treated with VEGF (50 ng/ml), and lysed after growth at 37oC for 2 min.图2：HUVE细胞经过VEGF刺激后，其裂解液中蛋白含量和试剂盒检测读值之间的关系。饥饿之后，用VEGF (50 ng/ml)在37°C 刺激HUVE细胞（铺至85%满）2分钟，然后裂解细胞。 |
