Biolegend, 904005, Alexa Fluor® 488 anti-RNA Polymerase II Antibody, 25μg

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 neuocleotide 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.
25μg
Verified Reactivity: Human, Mouse, Rat
Reported Reactivity: Other species
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: The immunogen used was a peptide containing 10 repeats of the synthetic peptide YSPTSPS using chemically synthesized phospho-ser5.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: ICC - Quality tested
Recommended Usage: Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/ml (1:500 dilution) is recommended. It is recommended that the reagent be titrated for optimal performance for each application. * Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses
Excitation Laser: Blue Laser (488 nm)
Application Notes: This antibody is effective in immunoblotting, immunoprecipitation and ELISA. It can also be used in chromatin immunoprecipitation assays. The antibody CTD4H8 recognizes the C-terminal repeat of the largest subunit of RNA polymerase II from HeLa and S. cerevisiae cells. This antibody recognizes both the phosphorylated and unphosphorylated forms of the RNA polymerase II. Based on sequence identify, this clone is liable to react with a broad range of species. Clone CTD4H8 is also known as 4H8 or CTD 4H8. Predicted MW is ~ 217 kD, Observed MW is on WB gel is ~240 kD
Application References(PubMed link indicates BioLegend citation): Kristjuhan A, et al. 2002. Mol. Cell. 10:925. Charos AE, et al. 2012. Genome Res. 22:1668. (ChIP) Rafalska-Metcalf IU, et al. 2010. PLoS One 5:e10272. (ICC) PubMed
RRID: AB_2728609 (BioLegend Cat. No. 904005) AB_2728610 (BioLegend Cat. No. 904006)
Biology Area: Cell Biology, Signal Transduction, Transcription Factors
Molecular Family: Nuclear Markers
Gene ID: 5430
UniProt: View information about RNA Polymerase II on UniProt.org
Clone: CTD4H8
Regulatory Status: RUO
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
Isotype: Mouse IgG1, κ