Rabbit POLR2A Monoclonal Antibody | anti-POLR2A antibody

Anti-Phospho-POLR2A (S5) Rabbit Monoclonal Antibody

Cat. Num. AAA1750248

• Gene Names: POLR2A; RPB1; RPO2; POLR2; POLRA; RPBh1; RPOL2; RpIILS; hsRPB1; hRPB220
• Reactivity: Human, Mouse, Rat
• Applications: Flow Cytometry, Functional Assay, Immunoprecipitation, Immunofluorescence, Immunohistochemistry, Immunocytochemistry, Western Blot
• Purity: Affinity-chromatography
• Synonyms: POLR2A; Monoclonal Antibody; Anti-Phospho-POLR2A (S5) Rabbit Monoclonal Antibody; DNA-directed RNA polymerase II subunit RPB1; RNA polymerase II subunit B1; 2.7.7.6; DNA-directed RNA polymerase II subunit A; DNA-directed RNA polymerase III largest subunit; RNA-directed RNA polymerase II subunit RPB1; 2.7.7.48; POLR2; anti-POLR2A antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat
• Clonality: Monoclonal
• Purity/Purification: Affinity-chromatography
• Form/Format: Liquid
• Sequence Length: 566

• Applicable Applications for anti-POLR2A antibody: Flow Cytometry (FC/FACS), Immunoprecipitation (IP), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB)
• Application Notes: WB: 1:500-1:2000; IHC: 1:50-1:200; ICC/IF: 1:50-1:200; IP: 1:20; FC/FACS: 1:40
• Immunogen: A synthesized peptide derived from human POLR2A
• Subcellular Localization: Nucleus.
• Contents: Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Preparation and Storage: Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Western blot analysis of Phospho-POLR2A (S5) expression in (1) HeLa cell lysate; (2) HeLa cell lysate treated with Lambda Phosphatase (MBS1750248).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-POLR2A monoclonal antibody overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for POLR2A )

Related Product Information for anti-POLR2A antibody

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 an 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.

NCBI and Uniprot Product Information

• NCBI GI #: 4505939
• NCBI GeneID: 5430
• NCBI Accession #: NP_000928.1
• NCBI GenBank Nucleotide #: NM_000937.4
• UniProt Accession #: P24928; Q6NX41; A6NN93; B9EH88
• Molecular Weight: 217176 MW
• NCBI Official Full Name: DNA-directed RNA polymerase II subunit RPB1
• NCBI Official Synonym Full Names: RNA polymerase II subunit A
• NCBI Official Symbol: POLR2A
• NCBI Official Synonym Symbols: RPB1; RPO2; POLR2; POLRA; RPBh1; RPOL2; RpIILS; hsRPB1; hRPB220
• NCBI Protein Information: DNA-directed RNA polymerase II subunit RPB1
• UniProt Protein Name: DNA-directed RNA polymerase II subunit RPB1
• UniProt Gene Name: POLR2A
• UniProt Synonym Gene Names: POLR2; RNA polymerase II subunit B1

NCBI Description

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, Jul 2008]

Uniprot Description

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. Regulation of gene expression levels depends on the balance between methylation and acetylation levels of tha CTD-lysines (). Initiation or early elongation steps of transcription of growth-factors-induced immediate early genes are regulated by the acetylation status of the CTD (PubMed:24207025). Methylation and dimethylation have a repressive effect on target genes expression ().