| 货号 | 7936C |
| 描述 | The PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-LAT (Tyr191). A LAT mouse antibody has been coated onto the microwells. After incubation with cell lysates, LAT protein (phosphorylated and non-phosphorylated) is captured by the coated antibody. Following extensive washing, a phospho-LAT (Tyr191) rabbit detection antibody is added to detect the captured phospho-LAT (Tyr191). HRP-linked anti-rabbit antibody is then used to recognize the bound detection antibody. The HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-LAT (Tyr191). 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-LAT (Tyr191) Sandwich ELISA Kit detects endogenous levels of Phospho-LAT when phosphorylated at Tyr191. As shown in Figure 1, a significant induction of LAT phosphorylation at Tyr191 can be detected in Jurkat cells following treatment with anti-CD3 using the Phospho-LAT (Tyr191) Sandwich ELISA Kit. The level of total LAT remains unchanged as shown by western analysis (Figure 1). 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 | LAT, a transmembrane adaptor protein expressed in T, NK and mast cells, is an important mediator for T cell receptor (TCR) signaling (1). Upon TCR engagement, activated Zap-70 phosphorylates LAT at multiple conserved tyrosine residues within SH2 binding motifs, exposing these motifs as the docking sites for downstream signaling targets (2,3). The phosphorylation of LAT at Tyr171 and Tyr191 enables the binding of Grb2, Gads/SLP-76, PLCγ1 and PI3 kinase through their SH2 domain and translocates them to the membrane. This process eventually leads to activation of the corresponding signaling pathways (1-4). |
| 存放说明 | 4C |
| 参考文献 | Wonerow, P. and Watson, S.P. (2001) Oncogene 20, 6273-6283. Zhang, W. et al. (1998) Cell 92, 83-92. Paz, P. E. et al. (2001) Biochem. J. 356, 461-471. Zhang, W. et al. (2000) J. Biol. Chem. 275, 23355-23361. |
Figure 1. Treatment of Jurkat cells with anti-CD3 stimulates phosphorylation of LAT at Tyr191 as detected by the PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Kit #7936, but does not affect the level of total LAT. Jurkat cells were starved for 48 hours and treated with 10 µg/ml anti-CD3 for 2 minutes at 37ºC. The absorbance readings at 450 nm are shown in the top figure, while the western blots, using LAT Antibody #9166 (left panel) or Phospho-LAT (Tyr191) Antibody #3584 (right panel), are shown in the bottom figure.图1.PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Kit #7936检测Jurkat细胞经anti-CD3诱导在Tyr191位点磷酸化的LAT, 但是不影响总的LAT蛋白。Jurkat细胞经过48小时的饥饿然后用10 µg/ml anti-CD3 在37ºC条件下处理 2 min。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 LAT Antibody #9166(左侧图)或Phospho-LAT (Tyr191) Antibody #3584(右侧图)。 | |
Figure 2. The relationship between lysate protein concentration from untreated and anti-CD3-treated Jurkat cells and the absorbance at 450 nm is shown.图2. 未经处理和经anti-CD3处理的Jurkat细胞的裂解液蛋白浓度与450 nm吸光度之间的关系。 |
