| 货号 | 7171C |
| 描述 | CSTs PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Zap-70 (Tyr319) 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, Phospho-Zap-70 (Tyr319) Ab is added to detect the captured phospho-Zap-70 (Tyr319) 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 phospho-Zap-70 (Tyr319) protein. * Antibodies in kit are custom formulations specific to kit.CST的 PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA Kit是固相夹心酶联免疫吸收剂检测试剂盒(ELISA)，它能够检测内源水平phospho-Zap-70 (Tyr319)蛋白。Zap-70鼠抗包裹到微孔的外表面。然后与细胞裂解液孵育，磷酸化和非磷酸化Zap-70蛋白都能够被抗体捕获。经过严格的冲洗后，Phospho-Zap-70 (Tyr319)抗体加入检测捕捉到的phospho-Zap-70 (Tyr319)蛋白。抗兔IgG, HRP连接抗体用于识别检测抗体的结合。HRP的底物TMB加入进行颜色反应。对反应的颜色，光密度的数值按比例换算成phospho-Zap-70 (Tyr319)蛋白的量。*试剂盒中的抗体是经过客户证实试剂盒特异的。 |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA Kit detects endogenous levels of phospho-Zap-70 (Tyr319) 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 total Zap-70 (phospho and non-phospho), 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).Syk 家族酪氨酸激酶Zap-70 在T 细胞和 NK 细胞中表达并在响应T细胞受体(TCR)而介导T细胞激活中发挥了重要的作用(1)。在TCR 招募下, Zap-70 通过自磷酸化和Src家族酪氨酸激酶Lck转磷酸化作用下快速的磷酸化几个酪氨酸残基(2-6)。酪氨酸的磷酸化与Zap-70激酶活性的增加和下游信号传导有关系。Zap-70 的表达与疾病的进程和慢性淋巴白血病患者的生存率有关(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 phosphorylation 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) Antibody #2701 (right panel) or Zap-70 Antibody #2705 (left panel), is shown in the bottom figure.图1：在Jurkat 细胞中，用过氧化氢处理磷酸化Zap-70的Tyr319位点, 并通过PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA kit #7171检测, 但是并未影响Zap-70 的总体水平（经PathScan® Total Zap-70 Sandwich ELISA kit #7172检测）。OD 450 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) Antibody #2701(右侧图) 或Zap-70 Antibody #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.图示为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试剂盒实验最佳密度读数的关系。在25ºC条件下，用过氧化氢(2 mM)处理Jurkat 细胞 (0.8 x 106 cells/ml) 2min，然后裂解。 |
