| 货号 | 87141S |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human\Mouse |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IP |
| 目标/特异性 | Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb recognizes endogenous levels of Btk protein only when phosphorylated at Tyr223. |
| 使用方法 | W IP |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | Brutons tyrosine kinase (Btk) is a member of the Btk/Tec family of cytoplasmic tyrosine kinases. Like other Btk family members, it contains a pleckstrin homology (PH) domain and Src homology SH3 and SH2 domains. Btk plays an important role in B cell development (1,2). Activation of B cells by various ligands is accompanied by Btk membrane translocation mediated by its PH domain binding to phosphatidylinositol-3,4,5-trisphosphate (3-5). The membrane-localized Btk is active and associated with transient phosphorylation of two tyrosine residues, Tyr551 and Tyr223. Tyr551 in the activation loop is transphosphorylated by the Src family tyrosine kinases, leading to autophosphorylation at Tyr223 within the SH3 domain, which is necessary for full activation (6,7). The activation of Btk is negatively regulated by PKCβ through phosphorylation of Btk at Ser180, which results in reduced membrane recruitment, transphosphorylation, and subsequent activation (8). The PKC inhibitory signal is likely to be a key determinant of the B cell receptor signaling threshold to maintain optimal Btk activity (8). |
| 存放说明 | -20C |
| 计算分子量 | 78 |
| 参考文献 | 1 . Khan, W.N. (2001) Immunol Res 23, 147-56. 2 . Lewis, C.M. et al. (2001) Curr Opin Immunol 13, 317-25. 3 . Salim, K. et al. (1996) EMBO J 15, 6241-50. 4 . Rameh, L.E. et al. (1997) J Biol Chem 272, 22059-66. 5 . Várnai, P. et al. (1999) J Biol Chem 274, 10983-9. 6 . Rawlings, D.J. et al. (1996) Science 271, 822-5. 7 . Park, H. et al. (1996) Immunity 4, 515-25. 8 . Kang, S.W. et al. (2001) EMBO J 20, 5692-702. |
Western blot analysis of extracts from Ramos cells, serum-starved overnight, then vehicle-treated (lane 1), treated with anti-human IgM (12 μg/ml for 10 min, lane 2), or pre-treated with Ibrutinib (1 μM for 60 min) prior to anti-IgM treatment (lane 3), using Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb (upper), and Btk (D3H5) Rabbit mAb #8547 (lower). | |
Immunoprecipitation of phosphorylated Btk (Tyr223) from Ramos cells serum-starved overnight, then treated with anti-IgM (12 ug/ml for 10 min), using Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb. Western blot analysis was performed using Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb. | |
Western blot analysis of extracts from Ramos cells, serum-starved overnight, then vehicle-treated (lane 1), treated with anti-human IgM (12 μg/ml for 10 min, lane 2), or treated with anti-human IgM followed by treatment with λ-phosphatase (lane 3), using Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb (upper), or Btk (D6H5) Rabbit mAb #8547 (lower). |
