| 货号 | 7172C |
| 描述 | CSTs PathScan® Total Zap-70 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Zap-70 protein. A Zap-70 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-Zap-70 proteins are captured by the coated antibody. Following extensive washing, Zap-70 Antibody is added to detect the captured phospho- and nonphospho-Zap-70 protein. Anti-rabbit 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 total Zap-70 protein. * Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total Zap-70 Sandwich ELISA Kit detects endogenous levels of Zap-70 enzyme. As shown in Figure 1, using this Sandwich ELISA Kit #7171, a significant induction of Phospho-Zap-70 (Tyr319) in Jurkat cells treated with hydrogen peroxide is detected. However, the level of Zap-70 (either untreated or treated), detected by the Total Zap-70 Sandwich ELISA Kit #7172, remains unchanged. 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 Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8). |
| 存放说明 | 4C |
| 参考文献 | Chu, D.H. et al. (1998) Immunol Rev 165, 167-80. Iwashima, M. et al. (1994) Science 263, 1136-9. Neumeister, E.N. et al. (1995) Mol Cell Biol 15, 3171-8. Chan, A.C. et al. (1995) EMBO J 14, 2499-508. Williams, B.L. et al. (1999) EMBO J 18, 1832-44. Di Bartolo, V. et al. (1999) J Biol Chem 274, 6285-94. Wiestner, A. et al. (2003) Blood 101, 4944-51. Crespo, M. et al. (2003) N Engl J Med 348, 1764-75. |
Figure 1: Treatment of Jurkat cells with hydrogen peroxide stimulates phopsphorylation of Zap-70 at Tyr319, detected by PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA kit #7171, but does not affect the level of total Zap-70 detected by PathScan® Total Zap-70 Sandwich ELISA kit #7172. OD 450 readings are shown in the top figure, while the corresponding Western blot using Phospho-Zap-70 (Tyr319) Ab #2701 (right panel) or Zap-70 Rabbit mAb #2705 (left panel), is shown in the bottom figure.图1: PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA kit #7171检测用过氧化氢处理Jurkat细胞诱导在Tyr319位点磷酸化Zap-70蛋白。但是经过PathScan® Total Zap-70 Sandwich ELISA kit #7172检测并未影响到Zap-70蛋白的总体水平。OD450的读数在图片的最上面显示,同时对应的Western免疫印迹在下面图片显示，所用抗体为Phospho-Zap-70 (Tyr319) Ab #2701(右侧图)或 Zap-70 Rabbit mAb #2705 (左侧图)。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and hydrogen peroxide treated Jurkat cells and kit assay optical density readings. Jurkat cells (0.8 x 106 cells/ml) were treated with hydrogen peroxide (2 mM) for 2 min at 25oC, and then lysed.图2: 未经处理和经过氧化氢处理的Jurkat细胞的裂解液蛋白浓度与吸光度之间的关系。Jurkat细胞(0.8 x 106 cells/ml) 在 25oC条件下经过氧化氢(2 mM) 处理2 min ，并裂解。 |
