| 货号 | 2629S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Mouse |
| 应用 | W/IP/ChIP |
| 使用方法 | WB(1:1000) IP (1:50) ChIP (1:50) |
| 供应商 | CST |
| 背景 | RNA polymerase II is a complex of 12 proteins that participates in transcription, mRNA processing, and transcription-coupled DNA repair (1,2). The largest subunit, Rpb1, contains a unique heptapeptide: Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7, which is repeated up to 52 times in the carboxy-terminal domain (CTD) of Rpb1 (3). This CTD is phosphorylated by cyclin-dependent kinases (CDKs), p44/42 MAPK, DNA-PKcs, and c-Abl (4). DNA damage caused by UV-light, hydrogen peroxide, or cisplatin results in ubiquitination and proteasomal degradation of Rpb1 (1,3). The kinase inhibitor 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole prevents ubiquitination of Rpb1, suggesting that CTD phosphorylation is required for proteolysis (5).RNA 聚合酶 II是由12种蛋白组成的复合体，参与转录、mRNA处理和转录偶联DNA修复(1,2)。最大的亚基Rpb1包含一个独特的七肽：Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7，在Rpb1的羧基末端结构域(CTD)发生高达52次的重复(3)。该结构域能够被周期蛋白依赖性激酶 (CDKs)，p44/42 MAPK，DNA-PKcs和c-Abl 磷酸化(4)。紫外线、过氧化氢和顺铂引起的DNA损伤能够导致Rpb1的泛素化和蛋白酶体降解(1,3)。激酶抑制剂5,6-二氯-1-β-D-呋喃核糖基苯并咪唑能够阻止Rpb1的泛素化表明CTD磷酸化为蛋白酶解所必需(5)。 |
| 存放说明 | -20C |
| 计算分子量 | 250 |
Western blot analysis of extracts from various cell lines using Rpb1 CTD (4H8) Mouse mAb.Western blot 方法检测不同细胞提取物，使用的抗体为Rpb1 CTD (4H8) Mouse mAb。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Rpb1 CTD (4H8) Mouse 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 SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human MYT-1 Exon 1 Primers #4493. 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.采用来自4 x 106 Hela细胞的交叉结合染色质进行免疫共沉淀实验，细胞通过紫外线处理（100 J/m2，恢复3小时）和10 μl Rpb1 CTD (4H8) Mouse mAb或者2 μl Normal Rabbit IgG #2729，使用的试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。随后通过实时PCR定量富集的DNA，使用的引物为SimpleChIP® Human RPL30 Exon 3 Primers #7014，SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490,和SimpleChIP® Human MYT-1 Exon 1 Primers #4493。每个样品的免疫沉淀DNA的量以相对于输入染色质总量（相当于1）的信号来表示 |
