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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| M, H | WB, ELISA, ChIP | R | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 04-1569 |
| Description | Anti-RNA polymerase II subunit B1 Antibody, clone 4F8 |
| Alternate Names |
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| Background Information | RNA polymerase II subunit B1 (RPB1) is the largest subunit of the RNA polymerase II complex. As a holoenzyme RNA polymerase II catalyzes transcription of eukaryotic DNA into RNA using the four ribonucleoside triphosphates as substrates. The RBP1 subunit, in combination with other polymerase subunits, forms a large central cleft that maintains contact between the active site of the enzyme, the DNA template, and the nascent RNA transcript. This subunit also contains a carboxy terminal domain (CTD) consisting of tandem heptapeptide repeats. In actively transcribing RNA polymerase ‘Ser-2’ and ‘Ser-5’ of the heptapeptide repeat are phosphorylated. Phosphorylation activates the RNA polymerase II beta subunit, allowing it to serve as an assembly platform for additional subunits that modulate initiation, elongation, termination and mRNA processing. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Purified rat monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mm NaCl) with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-RNA polymerase II subunit B1 Antibody, clone 4F8 is a Rat monoclonal antibody for detection of RNA polymerase II subunit B1 also known as DNA-directed RNA polymerase II A & has been validated in WB, ELISA. |
| Key Applications |
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| Application Notes | Chromatin Immunoprecipitation Analysis: A representative lot was used by independent laboraotry in ChIP. (Chapman, R., et Al. (2007). Science. 318(5857):1780 -1782.) |
| Biological Information | |
|---|---|
| Immunogen | Ovalbumin-conjugated linear peptide corresponding to human RNA polymerase II subunit B1. |
| Epitope | Unknown |
| Clone | 4F8 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rat |
| Specificity | This antibody recognizes RNA polymerase II subunit B1. |
| Isotype | IgG1κ |
| Species Reactivity |
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| Species Reactivity Note | Demonstrated to react with mouse. Predicted to react with human based on 100% sequence homology. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | 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]. |
| Gene Symbol |
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| Purification Method | Protein g Purified |
| UniProt Number |
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| UniProt Summary | 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. CATA;YTIC ACTIVITY: Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1). SUBUNIT STRUCTURE: Component of the RNA polymerase II (Pol II) complex consisting of 12 subunits. 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. Interacts (via C-terminus) with FTSJD2. 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 SIMILARITIES: Belongs to the RNA polymerase beta chain family. |
| Molecular Weight | ~ 220 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blot in NIH/3T3 cell lysates. Western Blot Analysis: 1 µg/mL of this antibody detected RNA polymerase II subunit B1 on 10 µg of NIH/3T3 cell lysates. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |