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merck millipore,默克密理博,17-672,ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set
产品名称：ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set
产品型号：17-672
This ChIPAb+ RNA Pol II -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
merck millipore,默克密理博,17-672,ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set
- 产品介绍
重要规格表
品种反应性 主要应用 Yeast (S. cerevisiae), H, M, R ChIP, WB, IHC, ICC 描述 产品目录编号 17-672 品牌系列 Upstate
商名 - Upstate
- ChIPAb+
描述 ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set 概述 All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ RNA Polymerase II (RNA Pol II) set includes the Anti-RNA Pol II, clone 8WG16 antibody, a negative control monoclonal antibody, and qPCR primers which amplify a 166 bp region within the promoter of the human GAPDH gene. The RNA Pol II clone 8WG16 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of RNA polymerase II-associated chromatin.Alternate Names - polymerase (RNA) II (DNA directed) polypeptide A, 220kDa
- DNA directed RNA polymerase II polypeptide A
- DNA-directed RNA polymerase II largest subunit, RNA<br />polymerase II 220 kd subunit
- DNA-directed RNA polymerase II subunit A
- DNA-directed RNA polymerase III largest subunit
- RNA polymerase II subunit B1
- polymerase (RNA) II (DNA directed) polypeptide A (220kD)
产品信息 格式 Ascites 控制 - Includes negative ascites and control primers specific for human GAPDH promoter.
演示 RNA Pol II (mouse monoclonal IgG2a, Clone 8WG16). One vial containing 50 μl ascites containing 0.05% sodium azide. Store at -20°C.
Negative Ascites. One vial containing 50 μl ascites containing 0.05% sodium azide. Store at -20°C.
Control Primers. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of GAPDH. Store at -20°C.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA
应用 应用 This ChIPAb+ RNA Pol II -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. 主要应用 - Chromatin Immunoprecipitation (ChIP)
- Western Blotting
- Immunohistochemistry
- Immunocytochemistry
应用说明 Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 μl of either Negative Ascites or anti-RNA Pol II clone 8WG16 antibody and the Magna ChIP G Kit (Part No. 17-611).
Successful immunoprecipitation of RNA Pol II associated DNA fragments was verified by qPCR using Control Primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
1:500-1:2000 dilution of a previous lot detected RNA Polymerase II in HeLa nuclear extract.
生物信息 免疫原品种 RNA Polymerase II purified from wheat germ extract. 表位 C-terminus of large subunit of Pol II 克隆 8WG16 宿主 Mouse 特异性 Recognizes RNA polymerase II, Mr ~220 kDa. 同种型 IgG2a 品种反应性 Yeast (S. cerevisiae)HumanMouseRat Species Reactivity Note Human, mouse, rat, yeast. Not tested in other species, but broad species reactivity is expected based on conservation of the C-terminal region of RNA pol II. 抗体类型 Monoclonal Antibody Entrez基因编号 - NM_000937.3
Entrez基因汇总 This gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. The product of this gene 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, this subunit, in combination with several other polymerase subunits, forms the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA. [provided by RefSeq] 基因符号 - hsRPB1
- POLR2
- POLRA
- RPB1
- RPBh1
- RPO2
- RPOL2
- RpIILS
- hRPB220
UniProt编号 - P24928
UniProt汇总 FUNCTION: DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.
Catalytic activity Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1).
SUBUNIT STRUCTURE: Component of the RNA polymerase II (Pol II) complex consisting of 12 subunits By similarity. The phosphorylated C-terminal domain interacts with FNBP3 and SYNCRIP. Interacts with SAFB/SAFB1. Interacts with CCNL1 and MYO1C By similarity. Interacts with CCNL2 and SFRS19. Component of a complex which is at least composed of HTATSF1/Tat-SF1, the P-TEFb complex components CDK9 and CCNT1, RNA polymerase II, SUPT5H, and NCL/nucleolin. Interacts with PAF1.
SUBCELLULAR LOCATION: Nucleus.
PTM: The tandem 7 residues repeats in the C-terminal domain (CTD) can be highly phosphorylated. The phosphorylation activates Pol II. Phosphorylation occurs mainly at residues 'Ser-2' and 'Ser-5' of the heptapepdtide repeat. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphataes, and a "CTD code" that specifies the position of Pol II within the transcription cycle has been proposed.
MISCELLANEOUS: The binding of ribonucleoside triphosphate to the RNA polymerase II transcribing complex probably involves a two-step mechanism. The initial binding seems to occur at the entry (E) site and involves a magnesium ion temporarily coordinated by three conserved aspartate residues of the two largest RNA Pol II subunits. The ribonucleoside triphosphate is transferred by a rotation to the nucelotide addition (A) site for pairing with the template DNA. The catalytic A site involves three conserved aspartate residues of the RNA Pol II largest subunit which permanently coordinate a second magnesium ion.
SEQUENCE SIMILARITY: Belongs to the RNA polymerase beta' chain family.
Contains 1 C2H2-type zinc finger.
产品使用声明 质量保证 Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µl of either a Negative Ascites or Anti- RNA Pol II, clone 8WG16 antibody and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation of RNA Pol II-associated DNA fragments was verified by qPCR using Control Primers (Please see figures).
Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
使用声明 - Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
储存和货运信息 存储条件 Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. 包装信息 数量 25 assays 包装 25 assays per set. Recommended use: ~2 μL antibody per chromatin immunoprecipitation (dependent upon biological context).
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