| 货号 | 7324C |
| 描述 | CSTs PathScan® Phospho-ALK (Tyr1604) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-ALK (Tyr1604) or phospho-NPM-ALK fusion protein. A Phospho-ALK (Tyr1604) Antibody has been coated onto the microwells. After incubation with cell lysates, only phospho-ALK or phospho-NPM-ALK proteins are captured by the coated antibody. Following extensive washing, an ALK Mouse mAb is added to detect the captured phospho-ALK or phospho-NPM-ALK fusion protein. Anti-mouse IgG, HRP-linked Antibody #7076 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of phospho-ALK (Tyr1604) or phospho-NPM-ALK proteins. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-ALK (Tyr1604) Sandwich ELISA Kit #7324 detects endogenous levels of phospho-ALK (Tyr1604) protein or phospho-NPM-ALK fusion protein. As shown in Figure 1, a high level of phosphorylated ALK (Tyr1604) protein or phospho-NPM-ALK fusion protein is detected in Karpas299 cells where ALK or NPM-ALK is constitutively phosphorylated. These high levels are abolished in Karpas299 cells lysed without addition of phosphatase inhibitors* to the lysis buffer. The levels of total ALK protein (phospho and nonphospho) detected by PathScan® Total ALK Sandwich ELISA Kit #7322 remain unchanged. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. * Phosphatase inhibitors includes sodium pyrophosphate, β-glycerophosphate and Na3VO4. |
| 供应商 | CST |
| 背景 | Anaplastic lymphoma kinase (ALK) is a tyrosine kinase receptor for pleiotrophin (PTN), a growth factor involved in embryonic brain development (1-3). In ALK-expressing cells, PTN induces phosphorylation of both ALK and the downstream effectors IRS-1, Shc, PLCγ, and PI3 kinase (1). ALK was originally discovered as an NPM (nucleophosmin)-ALK fusion protein produced by a translocation (4). The NPM-ALK fusion protein is a constitutively active, oncogenic tyrosine kinase associated with anaplastic lymphoma (4). Activation of PLCγ by NPM-ALK has been suggested to be a crucial step for its mitogenic activity and may be important in the pathogenesis of anaplastic lymphomas (5). A distinct ALK oncogenic fusion protein involving ALK and EML4 has been described from a non-small cell lung cancer cell line, with corresponding fusion transcripts present in some cases of lung adenocarcinoma. The short, amino-terminal region of the microtubule-associated protein EML4 is fused to the kinase domain of ALK (6,7). |
| 存放说明 | 4C |
| 参考文献 | Stoica, G.E. et al. (2001) J Biol Chem 276, 16772-9. Iwahara, T. et al. (1997) Oncogene 14, 439-49. Morris, S.W. et al. (1997) Oncogene 14, 2175-88. Morris, S.W. et al. (1994) Science 263, 1281-4. Bai, R.Y. et al. (1998) Mol Cell Biol 18, 6951-61. Rikova, K. et al. (2007) Cell 131, 1190-203. Takeuchi, K. et al. (2008) Clin Cancer Res 14, 6618-24. Soda, M. et al. (2007) Nature 448, 561-6. |
Figure 2: The relationship between protein concentration of phospho or non-phospho lysates and the absorbance at 450 nm is shown. Unstarved Karpas299 cells were cultured (106 cells/ml) and lysed with or without addition of phosphatase inhibitor to the lysis buffer (phospho or non-phospho lysate).图2：在波长450nm处对加入磷酸酶抑制剂处理（含磷酸化蛋白）和未经磷酸酶抑制剂处理的细胞裂解液（不含磷酸化蛋白）进行目的蛋白浓度检测的结果。Karpas299细胞在浓度为106 cells/ml时收集，并加入或不加磷酸酶抑制剂进行裂解处理。 |
