Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Rpb1 CTD (Ser2/5) Antibody #4735

DNA-directed RNA polymerase II largest subunit   POLR2A   RNA pol   RNA polymerase II 220 kd subunit   rnapII   RPB1  

No. Size Price
4735S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
4735 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 250 Rabbit
IHC-P 1:25

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin),


Species predicted to react based on 100% sequence homology: Hamster, S. cerevisiae,

Specificity / Sensitivity

Phospho-Rpb1 CTD (Ser2/5) Antibody detects endogenous levels of Rpb1 when the carboxy-termial domain heptapeptide repeat [Tyr1,Ser2,Pro3,Thr4,Ser5,Pro6,Ser7] is dually phosphorylated at Ser2 and Ser5 and singly phosphorylated at either Ser2 or Ser5.Phospho-Rpb1 CTD (Ser2/5) Antibody能够检测内源性羧基末端结构域七肽重复序列[Tyr1,Ser2,Pro3,Thr4,Ser5,Pro6,Ser7]丝氨酸(2,5位)双磷酸化和丝氨酸(2,5位)单磷酸化的Rpb1蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser2/5 of the Rpb1 CTD heptapeptide. Antibodies are purified by protein A and peptide affinity chromatography.该多克隆抗体是由合成的针对Rpb1蛋白CTD结构域七肽序列丝氨酸(2,5位)的磷酸化肽段免疫动物,采用蛋白A和多肽亲和层析技术纯化生产的。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-Rpb1 CTD (Ser2/5) Antibody.免疫组织化学方法检测石蜡包埋人类肺癌组织,显示核定位,使用的抗体为 Phospho-Rpb1 CTD (Ser2/5) Antibody。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Rpb1 CTD (Ser2/5) Antibody in the presence of control peptide (left) or antigen-specific peptide (right).免疫组织化学方法检测石蜡包埋人类结肠癌组织,使用的抗体为 Phospho-Rpb1 CTD (Ser2/5) Antibody,左图为与对照多肽共孵育,右图为抗原特异性多肽共孵育。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Rpb1 CTD (Ser2/5) Antibody.免疫组织化学方法检测石蜡包埋人类肾细胞癌组织,分为非处理组(左图)或λ磷酸酶处理组(右图),使用的抗体为 Phospho-Rpb1 CTD (Ser2/5) Antibody。

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF-7 cells, untreated or treated with doxorubicin (0.5 μM, 36 h), doxorubicin followed by λ Phosphatase NEB#P0753 (10,000 Units/ml, 1 h), nocodazole (50 ng/ml, 36 h) or nocodazole followed by λ Phosphatase, using Phospho-Rpb1 CTD (Ser2/5) Antibody (upper) or p53 Antibody #9282 (lower).Western blot方法检测MCF-7细胞,分为非处理组,阿霉素(0.5 μM)处理36小时组,λ磷酸酶(10,000 Units/ml,NEB#P0753)处理1小时后阿霉素处理组,诺考达唑(50 ng/ml)处理36小时组或λ磷酸酶处理后诺考达唑处理组,使用的抗体为Phospho-Rpb1 CTD (Ser2/5) Antibody (上图) 或 p53 Antibody #9282 (下图)。


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)。

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