| 货号 | 7953C |
| 反应种属 | Human/Dog |
| 应用 | ELISA |
| 目标/特异性 | The PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of Src when phosphorylated at Tyr416, as shown in Figure 1. This kit does not cross-react with the Src family members Yes, Lck, Lyn or Hck. Other family members have not been tested. 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 Src family of protein tyrosine kinases, which includes Src, Lyn, Fyn, Yes, Lck, Blk, and Hck, are important in the regulation of growth and differentiation of eukaryotic cells (1). Src activity is regulated by tyrosine phosphorylation at two sites, but with opposing effects. While phosphorylation at Tyr416 in the activation loop of the kinase domain upregulates enzyme activity, phosphorylation at Tyr527 in the carboxy-terminal tail by Csk renders the enzyme less active (2). |
| 存放说明 | 4C |
| 参考文献 | Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609. Hunter, T. (1987) Cell 49, 1-4. |
Figure 1. Phosphorylation of Src at Tyr416 in H202 stimulated MDCK cells (3 mM for 3 min) and untreated Calu-3 cells is detected by the PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit. In contrast, a low level of phospho-Src (Tyr416) is detected in untreated MDCK cells and Calu-3 cells lysed in the presence of λ-phosphatase. The absorbance readings at 450 nm are shown in the top figure while western blots, using Phospho-Src Family (Tyr416) (100F9) Rabbit mAb #2113 (upper) or Src (L4A1) Mouse mAb #2110 (lower), are shown in the bottom figure.图1：使用试剂盒PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit能检测H202刺激MDCK 细胞(3 mM 3 min)和未处理的Calu-3细胞中Src蛋白酪氨酸（416位点）的磷酸化。与此相反。用λ磷酸酶裂解未处理的MDCK，Calu-3细胞后只能检测到低水平的phospho-Src (Tyr416)。上图显示了450 nm处的光吸收读数，下图是western blots分析结果，使用的抗体是Phospho-Src Family (Tyr416) (100F9) Rabbit mAb #2113 (上侧)和Src (L4A1) Mouse mAb #2110 (下侧)。 | |
Figure 2. The relationship between lysate protein concentration from untreated and H202-treated MDCK cells and the absorbance at 450 nm is shown. MDCK cells were treated with H202 (3 mM) for 3 minutes at 37ºC and then lysed.图中显示了未经处理和H202处理的MDCK细胞溶解产物蛋白质浓度与450 nm 处光学密度读数之间的关系。MDCK细胞用H202 (3 mM)处理，37ºC 处理3min后裂解。 |
