| 货号 | 7176S |
| 描述 | CSTs PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-cdc2 (Tyr15) protein. A Phospho-cdc2 (Tyr15) Rabbit polyclonal Ab has been coated onto the microwells. After incubation with cell lysates, phospho-cdc2 (Tyr15) protein is captured by the coated antibody. Following extensive washing, cdc2 Mouse mAb is added to detect the captured phospho-cdc2 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 optical density for this developed color is proportional to the quantity of phospho-cdc2 (Tyr15) protein.CSTs PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Kit为固相三明治酶联免疫吸附试剂盒，能够检测内源性phospho-cdc2 (Tyr15)蛋白。Phospho-cdc2 (Tyr15) Rabbit polyclonal Ab包被于微孔上。与细胞裂解液孵育后，phospho-cdc2 (Tyr15)被包被抗体捕获。多次洗涤后，加入cdc2 Mouse mAb检测被捕获的phospho-cdc2蛋白。随后，用Anti-mouse IgG, HRP-linked Antibody识别结合的检测抗体。加入HRP底物(TMB)进行显色。显色过程中吸光度的强度和phospho-cdc2 (Tyr15)蛋白的量成正比。 |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Kit detects endogenous levels of phospho-cdc2 (Tyr15) protein. As shown in Figure 1, using the Phospho-cdc2 (Tyr15) ELISA Kit #7176, a significant induction of phospho-cdc2 (Tyr15) is detected in HeLa cells treated with Interleukin-4. 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 entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Thr14 and Tyr15 (2). Phosphorylation at Thr14 and Tyr15, resulting in inhibition of cdc2, can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).cdc2蛋白激酶的激活参与调节真核细胞进入有丝分裂，cdc2的激活受多个步骤的调控，包括周期蛋白的结合、cdc2苏氨酸161位的磷酸化(1)。然而，在细胞进入有丝分裂的过程中，激活cdc2的决定性步骤是苏氨酸14位和15位的去磷酸化(2)。Wee1 和 Myt1蛋白激酶通过磷酸化cdc2苏氨酸14位和15位，抑制cdc2活性(3,4)。而cdc25磷酸酯酶可能起到使cdc2苏氨酸14位和15位去磷酸化从而激活cdc2的作用(1,5)。 |
| 存放说明 | 4C |
| 参考文献 | Atherton-Fessler, S. et al. (1994) Mol Biol Cell 5, 989-1001. Norbury, C. et al. (1991) EMBO J 10, 3321-9. McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85. Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71. Hunter, T. (1995) Cell 80, 225-36. |
Figure 1: Treatment of HeLa cells with Interleukin-4 stimulates phosphorylation of cdc2 at Tyr15 as detected by PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA kit #7176, but does not affect the level of total cdc2 protein detected by cdc2 Antibody (#9116). OD 450 readings are shown in the top figure, while the corresponding Western blots using Phospho-cdc2 (Tyr15) Antibody #9111 (left panel) or cdc2 Antibody #9116 (right panel) is shown in the bottom figure.用IL-4处理HeLa细胞，使cdc2酪氨酸15位磷酸化，用PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA kit #7176检测，但并不影响cdc2总蛋白水平，cdc2总蛋白用cdc2 Antibody (#9116)检测。上图显示OD450读数，下图为相应的western blot分析，使用的抗体为Phospho-cdc2 (Tyr15) Antibody #9111（左图）和cdc2 Antibody #9116（右图）。 | |
Figure 2: The relationship between protein concentration of lysates from lambda phosphatase and IL-4 treated HeLa cells and kit assay optical density reading. HeLa cells (80% confluent) were treated with human IL-4 for 10 minutes (100 ng/ml).λ磷酸酶处理和IL-4处理的HeLa细胞裂解物蛋白浓度之间的关系，并分析光密度数据。人源IL-4 (100 ng/ml)处理HeLa细胞(80%的融合度)10分钟后收样。 |
