货号 | 7389S |
反应种属 | Human/Mouse/Rat/Monkey |
来源宿主 | Rabbit |
应用 | W/IP/IF-IC/ChIP |
目标/特异性 | CBP (D6C5) Rabbit mAb recognizes endogenous levels of total CBP protein. This antibody does not cross-react with p300 protein. |
使用方法 | WB(1:1000) IP (1:200) IF-IC (1:100) 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 p300 at Lys1499 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 |
Confocal immunofluorescent analysis of HeLa cells using CBP (D6C5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). 使用CBP (D6C5) Rabbit mAb 兔单抗(绿色)标记,共聚焦免疫荧光分析HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。蓝色= DRAQ5® #4084 (DNA荧光染料)。 | |
Western blot analysis of extracts from various cell lines using CBP (D6C5) Rabbit mAb. 使用CBP (D6C5) Rabbit mAb兔单抗,免疫印迹(Western blot)分析不同细胞的CBP蛋白水平。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells, treated with Forskolin #3828 (30 μM, 1h) and either 20 μl of CBP (D6C5) Rabbit mAb 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,来自Forskolin #3828 (30uM)处理过的4 x 106 293细胞的交联染色质以及20 µl CBP (D6C5) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用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。 |