| 货号 | 7939C |
| 描述 | The PathScan® Total FGFR2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of FGFR2 protein. An FGFR2 rabbit antibody has been coated on the microwells. After incubation with cell lysates, FGFR2 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a FGFR2 mouse detection antibody is added to detect captured FGFR2 protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total FGFR2 protein. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Total FGFR2 Sandwich ELISA Kit #7939 detects endogenous levels of FGFR2 protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 供应商 | CST |
| 背景 | Fibroblast growth factors (FGFs) produce mitogenic and angiogenic effects in target cells by signaling through cell surface receptor tyrosine kinases. There are four members of the FGF receptor family: FGFR-1 (flg), FGFR-2 (bek, KGFR), FGFR-3, and FGFR-4. Each receptor contains an extracellular ligand binding domain, a transmembrane domain, and a cytoplasmic kinase domain (1). Following ligand binding and dimerization, the receptors are phosphorylated at specific tyrosine residues (2). Seven tyrosine residues in the cytoplasmic tail of FGFR-1 can be phosphorylated: Tyr463, 583, 585, 653, 654, 730, and 766. Tyr653 and Tyr654 are important for catalytic activity of activated FGFR and are essential for signaling (3). The other phosphorylated tyrosine residues may provide docking sites for downstream signaling components such as Crk and PLCγ (4,5). |
| 存放说明 | 4C |
| 参考文献 | Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97. Reilly, J.F. et al. (2000) J Biol Chem 275, 7771-8. Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89. Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78. Larsson, H. et al. (1999) J Biol Chem 274, 25726-34. |
Figure 1. Constitutive phosphorylation of FGFR2 in KATO III cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-FGFR2 (panTyr) Sandwich ELISA Kit #7954 (upper, right). In contrast, a low level of phospho-FGFR2 protein is detected in KATO III cells lysed in the absence of phosphatase inhibitors (nonphospho lysate). Similar levels of total FGFR2 protein from both nonphospho and phospho lysates are detected by PathScan® Total FGFR2 Sandwich ELISA Kit #7939 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-FGF Receptor (Tyr653/654) Antibody #3471 (right) or a total FGFR2 Rabbit mAb (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and Na3VO4.图1：在磷酸酶抑制剂*存在时（磷酸化的裂解液），用PathScan® Phospho-FGFR2 (panTyr) Sandwich ELISA Kit #7954 能够检测到KATO III细胞中FGFR2的组成型表达（上，右）。相比之下，没有磷酸酶抑制剂*存在时（非磷酸化的裂解液），仅能检测到低水平的phospho- FGFR2的表达。用PathScan® Total FGFR2 Sandwich ELISA Kit #7939则能在磷酸化和非磷酸化的裂解液中检测到近似水平的总HER3/ErbB3的表达（上，左）。上图显示450nm处的吸光值，下图显示用Phospho-FGF Receptor (Tyr653/654) Antibody #3471（右）或total FGFR2 Rabbit mAb（左）进行免疫印迹检测的结果。*磷酸酶抑制剂中包含sodium pyrophosphate、β-glycerophosphate和 Na3VO4。 | |
Figure 2. The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. KATO III cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitors to the lysis buffer (phospho or nonphospho lysate, respectively).图2：磷酸化或非磷酸化的裂解中蛋白含量和450nm吸光值之间的关系。KATO III细胞在85%满时收样，并用加入或未加磷酸酶抑制剂的裂解液进行处理（磷酸化或非磷酸化）。 |
