| 货号 | 12070C |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-c-Abl (Tyr412) Sandwich ELISA Kit #12070 recognizes endogenous levels of Bcr-Abl or c-Abl protein when phosphorylated at Tyr412 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 | The c-Abl proto-oncogene encodes a nonreceptor protein tyrosine kinase that is ubiquitously expressed and highly conserved in metazoan evolution. c-Abl protein is distributed in both the nucleus and the cytoplasm of cells. It is implicated in regulating cell proliferation, differentiation, apoptosis, cell adhesion, and stress responses (1-3). c-Abl kinase activity is increased in vivo by diverse physiological stimuli including integrin activation; PDGF stimulation; and binding to c-Jun, Nck, and RFX1 (2,4). The in vivo mechanism for regulation of c-Abl kinase activity is not completely understood. Tyr245 is located in the linker region between the SH2 and catalytic domains. This positioning is conserved among Abl family members. Phosphorylation at Tyr245 is involved in the activation of c-Abl kinase (5). In addition, phosphorylation at Tyr412, which is located in the kinase activation loop of c-Abl, is required for kinase activity (6). |
| 存放说明 | 4C |
| 参考文献 | Wang, J.Y. et al. (2000) Oncogene 19, 5643-5650. |
Figure 2. The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. Unstarved K-562 cells were cultured (106 cells/ml) and lysed with or without addition of phosphatase inhibitors to the lysis buffer (phospho or nonphospho lysate, respectively).图2. 磷酸化及未磷酸化的胞裂解液蛋白浓度和450nm处吸光度的关系如图所示。未饥饿处理的K-562细胞培养(106 cells/ml)且被裂解，加入或不加入磷酸化抑制剂（磷酸化或磷酸化）。 | |
Figure 1. Constitutive phosphorylation of Bcr-Abl and c-Abl in K-562 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-c-Abl (Tyr412) Sandwich ELISA Kit #12070. In contrast, a low level of phospho-Bcr-Abl and phospho-c-Abl protein is detected in K-562 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Absorbance at 450 nm is shown in the top figure while corresponding western blots using c-Abl Antibody #2862 (left panel) and Phospho-c-Abl (Tyr412) (247C7) Rabbit mAb #2865 (right panel) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and Na3VO4.图1. K-562细胞裂解后持续磷酸化的Bcr-Abl和c-Abl在phosphatase inhibitors* (phospho lysate)存在的条件下，使用PathScan® Phospho-c-Abl (Tyr412) Sandwich ELISA Kit #12070进行检测。相对的，K-562细胞中低水平的phospho-Bcr-Abl和phospho-c-Abl在phosphatase inhibitors* (nonphospho lysate)存在的条件下也能被检测到。上图中展示了450nm处的吸光值，使用c-Abl抗体#2862（左）和Phospho-c-Abl (Tyr412) (247C7)兔mAb#2865（右）进行western blot分析的结果在下图中。 |
