- H14 (See other available formats)
- Regulatory Status
- Other Names
- DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II subunit B1, DNA-directed RNA polymerase II subunit A, RNA-directed RNA polymerase II subunit RPB1, DNA-directed RNA polymerase III largest subunit
- Mouse IgM, κ
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- Product Citations
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RPB1 is the catalytic and largest component of RNA polymerase II, which synthesizes mRNA precursors and many functional non-coding RNAs. It forms the polymerase active center together with RPB2, the second largest subunit. Polymerase II (Pol II) is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relatively 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 DNA template strand of the promoter is positioned within the central active site cleft of Pol II. Then, a bridging helix emanates from RPB1 and crosses the cleft near the catalytic site, which acts as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations during each nucleotide addition. This promotes translocation of Pol II. Pol II moves on the template during transcription elongation. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II's largest subunit (RPB1), which serves as a platform for assembling factors that regulate transcription initiation, elongation, termination, and mRNA processing. It can act as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, being able to conform as both a replicate and transcriptase for the viral RNA circular genome.Product Details
- Human, Mouse, Rat
- Antibody Type
- Host Species
- This antibody is provided in 50% glycerol in aqueous buffered solutions with preservatives.
- The antibody was purified by affinity chromatography and conjugated with HRP under optimal conditions.
- Lot-specific (please contact technical support for concentration and total µg amount, or use our Lookup tool if you have a lot number.)
- Storage & Handling
- Upon receipt, the antibody solution should be stored undiluted at -20°C, and protected from prolonged exposure to light.
WB - Quality tested
- Recommended Usage
Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.05 - 0.1 µg per ml (1:5000 - 1:10000 dilution). It is recommended that the reagent be titrated for optimal performance for each application.
25 µl and 100 µl of Direct-Blot™ HRP antibody can be used for approximately 5 and 20 Western blots, respectively, at the recommended concentration/dilution.
- Application Notes
Additional reported applications (for the relevant formats) include: immunoprecipitation2,3.
The clone H14 was developed against a purified phosphorylated form of RNA polymerase II extracted from a transformed cell line. Pol II has two physiologically important phosphorylation sites: ser-2 and ser-5. Both sites are found in the heptapeptide repeat YSPTSPS at the C-terminal domain. Clone H14 recognizes the phosphoserine 5 (ser-5) of Pol II. Clone H14 is useful for immunofluorescence and Western blotting.
This clone is not recommended for ChIP (Chromatin Immunoprecipitation) assays (as determined by in-house testing).
- Application References
- Brown JM, et al. 2008. J. Cell. Biol. 182:1083. (WB, IF)
- Bregman DB, et al. 1995. Cell Biol. 129:287-98. (IP)
- Ahn SH, et al. 2004. Mol. Cell. 13:67. (IP)
- Pellizzoni L, et al. 2001. Cell Biol. 152(1):75. (IF)
- Patturajan M, et al. 1998. J. Biol. Chem. 273(8):4689. (WB)
- Product Citations
AB_2734688 (BioLegend Cat. No. 920305)
AB_2734689 (BioLegend Cat. No. 920306)
- RPB1 contains a C-terminus composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain Ser and Thr residues that are phosphorylated in actively transcribing RNA polymerase. The predicted molecular weight is 220 kD.
Nucleus and cytosol.
- RPB1 is the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes.
- Biology Area
- Cell Biology, Cell Cycle/DNA Replication, Transcription Factors
- Molecular Family
- Nuclear Markers
- Gene ID
- 5430 View all products for this Gene ID
- View information about RNA Polymerase II RPB1 on UniProt.org
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