| 货号 | 39750 |
| 同种亚型 | IgG |
| 反应种属 | Human, Mouse, Other (Wide Range) |
| 来源宿主 | Rabbit |
| 应用 | ChIP/ChIP-Seq/Western Blot |
| 使用方法 | Validated Applications: ChIP: 4 - 10 µg per ChIP ChIP-Seq & ChIP-chip: 4 µl each WB: 0.5 - 2 µg/ml dilution |
| 供应商 | Active Motif | |
| 纯化方式 | Protein A Chromatography | |
| 免疫原 | This RNA pol II CTD phospho Ser5 antibody was raised against synthetic peptide containing the RNA Pol II heptad repeat consensus sequence phosphorylated at serine 5. | |
| 背景 | RNA pol II(RNA polymerase II) is responsible for synthesizing messenger RNA in eukaryotes. RNA pol II contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, RNA pol II, in combination with several other polymerase subunits, form the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA. During the transcription cycle, the CTD of the large subunit of RNA pol II is reversibly phosphorylated. RNA pol II containing unphosphorylated CTD is recruited to the promoter, whereas the hyperphosphorylated CTD form is involved in active transcription. Phosphorylation occurs at two sites within the heptapeptide repeat, at Serine 5 and Serine 2. RNA pol II Serine 5 phosphorylation is confined to promoter regions and is necessary for the initiation of transcription. | |
| 运输条件 | Blue Ice | |
| 存放说明 | Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage. | |
| 存储溶液 | Purified IgG in PBS (pH 7.5) with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an 计算分子量 | 210 kDa |
| 参考文献 |
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RNA pol II CTD phospho Ser5 antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 2.3 million HL-60 cells and 4 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 26 million sequence tags were mapped to identify RNA pol II phospho Ser5 binding. The image shows a 1.2 million base pair region on chromosome 15 with the expected enrichment at gene promoters. | |
RNA pol II CTD phospho Ser5 antibody tested by ChIP analysis. Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 µg of RNA pol II CTD phospho Ser5 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the PABPC1 gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG (arbitrarily assigned a value of 1) using the ddCT method. | |
RNA pol II CTD phospho Ser5 antibody tested by Western blot. Nuclear extract of HeLa cells (20 µg) probed with RNA pol II CTD phospho Ser5 antibody at a dilution of 0.5 µg/ml. | |
RNA pol II CTD phospho Ser5 pAb tested by dot blot analysis. Dot blot analysis was used to confirm the specificity of RNA pol II CTD phospho Ser5 antibody for phospho-Ser5 of the RNA Pol II C-terminal domain heptad repeat. Modified and unmodified peptides were spotted onto PVDF and probed with the antibody at a dilution of 0.2 µg/ml. Decreasing amounts of peptide were spotted in each row. Lane 1: Peptide phosphorylated at CTD repeat serine 2. Lane 2: Unmodified CTD repeat serine 2 peptide. Lane 3: Peptide phosphorylated at CTD repeat serine 5. Lane 4: Unmodified CTD repeat serine 5 peptide. |
