| 货号 | 7903C |
| 描述 | PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Bcr-Abl and c-Abl proteins. A c-Abl rabbit antibody has been coated on the microwells. After incubation with cell lysates, Bcr-Abl and c-Abl protein (phospho and nonphospho) are captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse detection antibody is added to detect captured tyrosine-phosphorylated Bcr-Abl and c-Abl 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 tyrosine-phosphorylated Bcr-Abl and c-Abl protein. Antibodies in kit are custom formulations specific to kit.PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit是固相夹心酶联免疫吸附试验（ELISA），能够检测内源性水平的酪氨酸磷酸化Bcr-Abl和c-Abl蛋白。c-Abl兔抗包被微孔板。细胞裂解液孵育后，Bcr-Abl和c-Abl（磷酸和非磷酸）蛋白由包被的抗体捕获。充分冲洗后，加入磷酸酪氨酸鼠检测抗体检测捕获的酪氨酸磷酸化的Bcr-Abl和c-Abl蛋白。随后用抗鼠IgG，HRP-标记的抗体识别结合的检测蛋白。加入HRP底物TMB进行显色。显色的吸光度值与酪氨酸磷酸化的Bcr-Abl和c-Abl蛋白的量成正比。试剂盒中的抗体是针对试剂盒定制的。 |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit #7903 detects endogenous levels of Bcr-Abl or c-Abl protein when phosphorylated at Tyr residues 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 Bcr gene was orginally identified by its presence in the chimeric Bcr-Abl oncogene (1). The amino-terminal region of Bcr contains an oligomerization domain, a serine/threonine kinase domain, and a region that binds SH2 domains. The middle of the protein has a PH domain and a region of sequence similarity to the guanine nucleotide exchange factors for the Rho family of GTP binding proteins. The carboxy-terminal region may be involved in a GTPase activating function for the small GTP-binding protein Rac (2,3). The function of wild type Bcr in cells remains unclear. PDGF receptor may use Bcr as a downstream signaling mediator (4). Research studies have shown that the Bcr-Abl fusion results in production of a constitutively active tyrosine kinase, which causes chronic myelogenous leukemia (CML) (5). Tyr177 of Bcr is phosphorylated in the Bcr-Abl fusion protein, which plays an important role in transforming the activity of Bcr-Abl (6). Phosphorylated Tyr177 provides a docking site for Gab2 and GRB2 (7,8). |
| 存放说明 | 4C |
| 参考文献 | Wang, J.Y. et al. (2000) Oncogene 19, 5643-5650. Van Etten, R.A. et al. (1999) Trends Cell. Biol. 9, 179-182. Danial, N.N. et al. (2000) Oncogene 19, 2523-2531. Shaul, Y. et al. (2000) Cell Death Differ. 7, 10-16. Brasher, B.B. et al. (2000) J. Biol. Chem. 275, 35631-35637. Pluk, H. et al. (2002) Cell 108, 247-259. |
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 (panTyr) Sandwich ELISA Kit #7903. 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) and Phospho-c-Abl (Tyr412) (247C7) Rabbit mAb #2865 (right) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and Na3VO4. 图1. 在磷酸酶抑制剂*（磷酸裂解物）存在的情况下裂解K-562细胞，PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit #7903检测到组成型磷酸化的Bcr-Abl和c-Abl。相反，在磷酸酶抑制剂*（非磷酸裂解物）缺乏时裂解K-562细胞，可以检测到低水平的磷酸化Bcr-Abl和磷酸化c-Abl蛋白。上图显示450 nm处的吸光度值，而对应的western blots结果见下图，western blots实验使用的抗体为c-Abl Antibody #2862 (左图) 和Phospho-c-Abl (Tyr412) (247C7) Rabbit mAb #2865 (右图)。*磷酸抑制剂包含焦磷酸钠、β-磷酸甘油和Na3VO4. | |
Figure 2. The relationship between protein concentration of phospho or non-phospho 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 inhibitor to the lysis buffer (phospho or non-phospho lysate, respectively). 图2.如图所示，磷酸或非磷酸化裂解物的蛋白浓度和450 nm处吸光度值的关系。未饥饿的K-562细胞（106 个细胞/ml）经培养后，用含或不含磷酸酶抑制剂的裂解液（分别为磷酸或非磷酸裂解物）进行裂解。 |
