| 货号 | 2330T |
| 目标/特异性 | Each antibody in the Raf Family Antibody Sampler Kit recognizes only its specific target and does not cross react with other Raf family members. |
| 供应商 | CST |
| 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 |
| 参考文献 | Avruch, J. et al. (1994) Trends Biochem Sci 19, 279-83. Chong, H. et al. (2001) EMBO J 20, 3716-27. King, A.J. et al. (1998) Nature 396, 180-3. Fabian, J.R. et al. (1993) Mol Cell Biol 13, 7170-9. Mason, C.S. et al. (1999) EMBO J 18, 2137-48. Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4. Sprenkle, A.B. et al. (1997) FEBS Lett 403, 254-8. Marais, R. et al. (1997) J Biol Chem 272, 4378-83. Guan, K.L. et al. (2000) J Biol Chem 275, 27354-9. Davies, H. et al. (2002) Nature 417, 949-54. Dougherty, M.K. et al. (2005) Mol Cell 17, 215-24. |
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.) | |
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition. | |
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition. | |
Western blot analysis of extracts from HT-29, NIH-3T3 and C6 cell lysates, using A-Raf Antibody. | |
Western blot analysis of extracts from NIH3T3, HeLa and COS cells, untreated or treated with TPA, using Phospho-c-Raf (Ser338) (56A6) Rabbit mAb. | |
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 analysis of extracts from Raji and HeLa cells, untreated or TPA-treated (30 minutes), using Phospho-A-Raf (Ser299) Antibody (upper) or A-Raf Antibody, #4432 (lower). | |
Western blot analysis of extracts from Raji and HeLa cells treated with TPA (200nM, 30 minutes) using Phospho-B-Raf (Ser445) Antibody (top) and total B-Raf Antibody (bottom). | |
Western blot analysis of extracts from untreated or TPA-treated 293 and NIH/3T3 cells, using Phospho-c-Raf (Ser289/296/301) Antibody. | |
Western blot analysis of extracts from HeLa, C2C12 and NBT-II cells, using B-Raf (55C6) Rabbit mAb. | |
Western blot analysis of extracts from Raji and HeLa cells, untreated or TPA-treated (200 nM, 30 minutes) using Phospho-B-Raf (Ser445) Antibody #2696 (top) and total B-Raf Antibody (bottom).Western blot方法检测细胞提取物：未经处理和TPA处理30min的Raji，HeLa细胞，使用的抗体是Phospho-A-Raf (Ser299) Antibody #4431 (上图)和A-Raf Antibody #4432 (下图)。 | |
Western blot analysis of extracts from Raji and HeLa cells, untreated or TPA-treated (30 minutes), using Phospho-A-Raf (Ser299) Antibody #4431 (upper) or A-Raf Antibody #4432 (lower). | |
Western blot analysis of extracts from HT29, NIH/3T3 and C6 cell lysates using A-Raf Antibody #4432.Western blot方法检测细胞提取物：HT29, NIH/3T3，C6细胞，使用的抗体是A-Raf Antibody #4432。 | |
Western blot analysis of extracts from HeLa cells, untreated or TPA-treated, using Phospho-c-Raf (Ser259) Antibody #9421 (upper), or a total c-Raf antibody (lower).Western blot方法检测细胞提取物：未经处理、TPA处理的HeLa细胞，使用的抗体是Phospho-c-Raf (Ser259) Antibody #9421 (上图), a total c-Raf antibody (下图)。 | |
Western blot analysis of extracts from 293 cells, untreated or TPA-treated (200 nM for 30 minutes), using c-Raf Antibody #9422 (lower) or Phospho-c-Raf (Ser338) Rabbit mAb #9427 (upper).Western blot方法检测细胞提取物：未经处理、TPA(200 nM 30 min)处理的293细胞，使用的抗体是c-Raf Antibody #9422 (下图)和Phospho-c-Raf (Ser338) Rabbit mAb #9427 (上图)。 |
