| 货号 | 9421T |
| 反应种属 | Human,Mouse,Rat,Monkey,Xenopus, |
| 来源宿主 | Rabbit |
| 应用 | WB, IP |
| 目标/特异性 | Phospho-c-Raf (Ser259) Antibody detects endogenous levels of c-Raf only when phosphorylated at Ser259. |
| 使用方法 | WB(1:1000) IP (1:50) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | A-Raf, B-Raf and c-Raf (Raf-1) are the main effectors recruited by GTP-bound Ras to activate the MEK-MAP kinase pathway (1). Activation of c-Raf is the best understood and involves phosphorylation at multiple activating sites including Ser338, Tyr341, Thr491, Ser494, Ser497 and Ser499 (2). p21-activated protein kinase (PAK) has been shown to phosphorylate c-Raf at Ser338 and the Src family phosphorylates Tyr341 to induce c-Raf activity (3,4). Ser338 of c-Raf corresponds to similar sites in A-Raf (Ser299) and B-Raf (Ser445), although this site is constitutively phosphorylated in B-Raf (5). Inhibitory 14-3-3 binding sites on c-Raf (Ser259 and Ser621) can be phosphorylated by Akt and AMPK, respectively (6,7). While A-Raf, B-Raf and c-Raf are similar in sequence and function, differential regulation has been observed (8). Of particular interest, B-Raf contains three consensus Akt phosphorylation sites (Ser364, Ser428 and Thr439) and lacks a site equivalent to Tyr341 of c-Raf (8,9). The B-Raf mutation V600E results in elevated kinase activity and is commonly found in malignant melanoma (10). Six residues of c-Raf (Ser29, Ser43, Ser289, Ser296, Ser301 and Ser642) become hyperphosphorylated in a manner consistent with c-Raf inactivation. The hyperphosphorylation of these six sites is dependent on downstream MEK signaling and renders c-Raf unresponsive to subsequent activation events (11). |
| 存放说明 | -20C |
| 计算分子量 | 74 |
| 参考文献 | 1 . Avruch, J. et al. (1994) Trends Biochem Sci 19, 279-83. 2 . Chong, H. et al. (2001) EMBO J 20, 3716-27. 3 . King, A.J. et al. (1998) Nature 396, 180-3. 4 . Fabian, J.R. et al. (1993) Mol Cell Biol 13, 7170-9. 5 . Mason, C.S. et al. (1999) EMBO J 18, 2137-48. 6 . Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4. 7 . Sprenkle, A.B. et al. (1997) FEBS Lett 403, 254-8. 8 . Marais, R. et al. (1997) J Biol Chem 272, 4378-83. 9 . Guan, K.L. et al. (2000) J Biol Chem 275, 27354-9. 10 . Davies, H. et al. (2002) Nature 417, 949-54. 11 . Dougherty, M.K. et al. (2005) Mol Cell 17, 215-24. |
Western blot analysis of extracts from HeLa cells, untreated or TPA-treated, using Phospho-c-Raf (Ser259) Antibody (upper), or a total c-Raf antibody (lower).Western blot方法检测细胞提取物：未经处理和TPA处理的HeLa细胞。使用的抗体是Phospho-c-Raf (Ser259) Antibody (上图)和a total c-Raf antibody (下图)。 | |
Site specificity of Phospho-c-Raf (Ser259) Antibody: Western blot analysis of recombinant Myc-tagged c-Raf protein, wild-type (lanes 1 and 3) and S259A mutant (lanes 2 and 4), using Phospho-Raf (Ser259) Antibody or a Myc antibody. (Provided by Dr. Guri Tzivion, Massachusetts General Hospital.)Phospho-c-Raf (Ser259) Antibody位点特异性：Western blot方法检测Myc标记的c-Raf重组蛋白，野生型(1、3号条带)和S259A 突变体(2、4号条带)。使用的抗体是Phospho-Raf (Ser259) Antibody 和Myc antibody。（由Dr. Guri Tzivion提供, Massachusetts General Hospital)。 |
