| 货号 | 7970C |
| 描述 | The PathScan® Phospho-Syk (Tyr525/526) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Syk when phosphorylated at tyrosine 525/526. A phospho-Syk (Tyr525/526) Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Syk phosphorylated at Tyrosine525/526 is captured by the coated antibody. Following extensive washing, a Syk mouse monoclonal detection antibody is added to detect the captured Syk protein. HRP-linked Anti-mouse Ab is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Syk phosphorylated on Tyr525/526. 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-Syk (Tyr525/526) Sandwich ELISA Kit #7970 detects endogenous levels of Syk protein when phosphorylated at Tyr525/526. 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 | Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells, and Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation of Tyr323 provides a direct binding site to the TKB domain of Cbl (6,7). Tyrosine 352 of Syk is involved in the association of PLC-γ1 (8). Tyrosines 525 and 526 are located in the activation loop of the Syk kinase domain, and phosphorylation of Tyr525/526 of human Syk (equivalent to the Tyr519/520 of mouse Syk) is essential for Syk function (9). |
| 存放说明 | 4C |
| 参考文献 | Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80. Turner, M. et al. (2000) Immunol Today 21, 148-54. Coopman, P.J. et al. (2000) Nature 406, 742-7. Deckert, M. et al. (1998) J Biol Chem 273, 8867-74. Rao, N. et al. (2001) EMBO J 20, 7085-95. Law, C.L. et al. (1996) Mol Cell Biol 16, 1305-15. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7. |
Figure 1. Treatment of Ramos cells with goat anti-human IgM stimulates tyrosine phosphorylation of Syk, detected by PathScan® Phospho-Syk (Tyr525/526) Sandwich ELISA Kit #7970, but does not affect the levels of total Syk protein detected by western blot analysis. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots, using Syk Antibody #2712 (left panel), or Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb #2710 (right panel) are shown in the bottom figure.图1.在Ramos细胞中，用goat anti-human IgM刺激产生酪氨酸位点磷酸化得Syk,并通过PathScan® Phospho-Syk (Tyr525/526) Sandwich ELISA Kit #7970检测, 经western免疫印迹检测表明总Syk蛋白水平并未受到影响 。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Syk Antibody #2712(左侧图) 或Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb #2710 (右侧图)。 | |
Figure 2. The relationship between protein concentration of untreated or goat anti-human IgM treated Ramos cell lysates and the absorbance at 450 nm is shown. Cells were serum starved overnight and then treated with goat anti-human IgM (10 μg/ml) for 10 min. at 37ºC.图2.图示为未经处理和经goat anti-human IgM处理的Ramos细胞的裂解液蛋白浓度与450 nm 吸收峰的关系，在37ºC条件下，细胞经过夜血清饥饿并经goat anti-human IgM (10 μg/ml)处理10 min。 |
