| 货号 | 7073C |
| 描述 | The PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IKKα when phosphorylated at Ser176/180. A phospho-IKKα (Ser176/180) rabbit mAb has been coated onto the microwells. After incubation with cell lysates, phospho-IKKα (Ser176/180) protein is captured by the coated antibody. Following extensive washing, an IKKα mouse detection mAb is added to detect the captured IKKα protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of IKKα phosphorylated at Ser176/180. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | The PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of IKKα protein when phosphorylated at Ser176/180, as shown in Figure 1. 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 NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation; Ser177 and Ser181 in the activation loop of IKKβ (serine 176 and 180 in IKKα) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13). |
| 存放说明 | 4C and -20C |
| 参考文献 | Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8. Brown, K. et al. (1995) Science 267, 1485-8. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83. Chen, Z.J. et al. (1996) Cell 84, 853-62. Zandi, E. et al. (1997) Cell 91, 243-52. Karin, M. (1999) Oncogene 18, 6867-74. DiDonato, J.A. et al. (1997) Nature 388, 548-54. Mercurio, F. et al. (1997) Science 278, 860-6. Johnson, L.N. et al. (1996) Cell 85, 149-58. Delhase, M. et al. (1999) Science 284, 309-13. |
Figure 1. Treatment of differentiated THP-1 cells with LPS stimulates phosphorylation of IKKα at Ser176/180 as detected by the PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit #7073 but does not affect the levels of total IKKα detected by PathScan® Total IKKα Sandwich ELISA Kit #7078. Differentiated THP-1 cells were treated with 1 μg/ml LPS for 10 minutes. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using IKKα Antibody #2682 (left) or Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697 (right) are shown in the bottom figure.图1.用LPS(1 μg/ml ,10 min)处理经过分化的THP-1细胞，刺激IKKα在Ser176/180位点的磷酸化,并用PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit #7073检测, 但是通过PathScan® Total IKKα Sandwich ELISA Kit #7078检测并未影响总IKKα蛋白。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IKKα Antibody #2682(左图) 或Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697 (右图)。 | |
Figure 2. The relationship between the protein concentration of lysates from untreated and LPS-treated THP-1 cells and the absorbance at 450 nm using the PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit is shown.图2.图示为未经处理和经LPS处理的THP-1细胞的裂解液蛋白浓度与试剂盒PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit最佳450 nm读数的关系。 |
