| 货号 | 4771S |
| 反应种属 | Human/Mouse/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/ChIP |
| 目标/特异性 | Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody detects endogenous levels of CBP or p300 only when acetylated at lysine 1535 or lysine 1499, respectively. |
| 使用方法 | WB(1:1000) IP (1:50) ChIP (1:25) |
| 供应商 | CST |
| 背景 | CBP (CREB-binding protein) and p300 are highly conserved and functionally related transcriptional co-activators that associate with transcriptional regulators and signaling molecules, integrating multiple signal transduction pathways with the transcriptional machinery (1,2). CBP/p300 also contain histone acetyltransferase (HAT) activity, allowing them to acetylate histones and other proteins (2). Phosphorylation of p300 at Ser89 by PKC represses its transciptional acitivity, and phosphorylation at the same site by AMPK disrupts the association of p300 with nuclear receptors (3,4). Ser1834 phosphorylation of p300 by Akt disrupts its association with C/EBPβ (5). Growth factors induce phosphorylation of CBP at Ser437, which is required for CBP recruitment to the transcription complex (6). CaM kinase IV phosphorylates CBP at Ser302, which is required for CBP-dependent transcriptional activation in the CNS (7). The role of acetylation of CBP/p300 is of particular interest (2,8). Acetylation of Lys1499 of p300 has been demonstrated to enhance its HAT activity and affect a wide variety of signaling events (9). |
| 存放说明 | -20C |
| 计算分子量 | 300 |
Western blot analysis of hypo- or hyper-acetylated recombinant p300 HAT domains, either wild-type or K1499R mutant, using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody (upper). Also shown in the corresponding coomassie stained SDS-PAGE gel (lower). (Details are described in Thompson, P.A. et al. (2004) Nat. Struct. Mol. Biol. 11, 308-315.) 使用Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody (上图)，免疫印迹(Western blot)分析hypo- or hyper-乙酰化的重组p300 HAT结构域，wild-type 或K1499R突变。 下图是相应的考马斯亮蓝染色的SDS-PAGE胶的结果。(Details are described in Thompson, P.A. et al. (2004) Nat. Struct. Mol. Biol. 11, 308-315.) | |
Western blot analysis of extracts from A431, NIH/3T3, COS and PC12 cells, using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody. 使用Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody，免疫印迹(Western blot)分析A431、NIH/3T3、COS和PC12细胞中Acetyl-CBP (Lys1535)/p300 (Lys1499)的蛋白水平。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells treated with Forskolin #3828 (30uM) and either 20 μl of Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. 使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，用Forskolin #3828 (30uM)处理过的4 x 106 293细胞的交联染色质以及20 µl Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829和SimpleChIP® Human α Satellite Repeat Primers #4486，浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号，这相当于一。 |
