Product Details

Uniprot No.

Target Names

Alternative Names

DNA directed RNA polymerase II A antibody; DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody; DNA-directed RNA polymerase II subunit A antibody; DNA-directed RNA polymerase II subunit RPB1 antibody; DNA-directed RNA polymerase III largest subunit antibody; hRPB220 antibody; hsRPB1 antibody; POLR2 antibody; Polr2a antibody; POLRA antibody; Polymerase (RNA) II (DNA directed) polypeptide A 220kDa antibody; Polymerase (RNA) II (DNA directed) polypeptide A antibody; RNA polymerase II subunit B1 antibody; RNA-directed RNA polymerase II subunit RPB1 antibody; RPB1 antibody; RPB1_HUMAN antibody; RPBh1 antibody; RpIILS antibody; RPO2 antibody; RPOL2 antibody

Species Reactivity

Human

Immunogen

A synthesized peptide derived from Human Phospho-POLR2A (S5)

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

2H4

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, IF

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
IF	1:20-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

RNA polymerase II, the enzyme responsible for transcribing protein-coding genes into mRNA, is regulated through dynamic phosphorylation of its largest subunit, POLR2A. Phosphorylation at serine 5 within the C-terminal domain heptad repeats serves as a critical marker of transcription initiation, recruiting capping enzymes and coordinating the transition from promoter clearance to productive elongation. Detecting this specific phosphorylation event provides researchers with a window into active transcriptional states and the regulatory mechanisms governing gene expression programs.

This recombinant monoclonal antibody, generated against a synthetic phosphopeptide corresponding to the Ser5-phosphorylated region of human POLR2A, offers the reproducibility essential for phospho-specific detection. Because recombinant production ensures a sequence-defined binding site and eliminates the lot-to-lot variability inherent in traditional hybridoma methods, researchers can confidently compare results across extended studies or between laboratories.

Validation across multiple experimental platforms demonstrates this antibody's versatility in your workflow. Western blot analysis detects a band at the expected 270 kDa molecular weight in HeLa, A549, and 293 whole cell lysates, confirming specificity for the full-length phosphorylated protein. Immunocytochemistry performed on EGF-stimulated HeLa cells reveals the expected nuclear localization pattern, while immunofluorescence staining with DAPI counterstaining provides clear visualization of phospho-POLR2A distribution within the nuclear compartment.

Whether investigating transcriptional regulation, chromatin dynamics, or signal-dependent gene activation, this antibody delivers reliable phospho-specific detection of a fundamental epigenetic marker. Its compatibility with western blotting, immunohistochemistry, and immunofluorescence applications supports diverse experimental approaches in transcription and nuclear signaling research.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Target Background

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. 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. Methylation and dimethylation have a repressive effect on target genes expression.; (Microbial infection) 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.

Gene References into Functions

XPC is an RNA polymerase II cofactor recruiting ATAC coactivator complex to promoters by interacting with E2F1. PMID: 29973595
weak, multivalent interactions between TAF15 fibrils and heptads throughout RNA pol II CTD collectively mediate complex formation. PMID: 28945358
This shows that CDK9 stimulates release of paused polymerase and activates transcription by increasing the number of transcribing polymerases and thus the amount of mRNA synthesized per time. PMID: 28994650
Results identified rs2071504 in POLR2A gene to be associated with poor overall and disease-free survival of patients with an early-stage non-small cell lung cancer. PMID: 28922562
Dara indicate that hydrogen peroxide alters RNA polymerase II (Pol II) occupancy at promoters and enhancers genome-wide. PMID: 28977633
Rpb1/2 dynamics help govern the decision between sense and divergent antisense transcription. PMID: 28506463
The results showed heterogeneity in the responses of individual KSHV episomes to stimuli within a single reactivating cell; those episomes that did respond to stimulation, aggregated within large domains that appear to function as viral transcription factories. A significant portion of cellular RNA polymerase II was trapped in these factories and served to transcribe viral genomes. PMID: 28331082
Data show that inhibition of VCP/p97, or siRNA-mediated ablation of VCP/p97 impairs ultraviolet radiation (UVR)-induced RNA polymerase II (RNAPII) degradation. PMID: 28036256
Role of chromatin-bound EGFR and ERK kinases in RNA polymerase 2 transcription PMID: 27587583
recurrent somatic mutations in POLR2A hijack this essential enzyme and drive meningioma neoplasia PMID: 27548314
the Elongin A ubiquitin ligase and the CSB protein function together in a common pathway in response to Pol II stalling and DNA damage PMID: 28292928
By studying global gene expression patterns and genome-wide DNA-binding patterns of CGGBP1, it has been shown that a possible mechanism through which it affects the expression of RNA Pol II-transcribed genes in trans depends on Alu RNA. PMID: 25483050
Using a 7,781-sample pan-cancer dataset, we first confirmed this in POLR2A are known to confer elevated sensitivity to pharmacological suppression.hese include the POLR2A interacting protein INTS10 as well as genes involved in mRNA splicing, nonsense-mediated mRNA PMID: 28027311
HIV Tat precisely controls RNA polymerase II recruitment and pause release to fine-tune the initiation and elongation steps in target genes. PMID: 26488441
TOP1 bound at promoters was discovered to become fully active only after pause-release. This transition coupled the phosphorylation of the carboxyl-terminal-domain (CTD) of RNA polymerase II (RNAPII) with stimulation of TOP1 above its basal rate, enhancing its processivity. PMID: 27058666
Its variant is not related to sporadic PD in Chinese Han population. PMID: 26432391
Data suggest RNA polymerase II (POLR2A) is extensively modified on its unique C-terminal domain (CTD) by O-GlcNAc transferase (OGT); efficient O-GlcNAcylation requires a minimum of 20 heptad CTD repeats in POLR2A and more than half of NTD of OGT. PMID: 26807597
Serine phosphorylation stimulates whereas tyrosine phosphorylation inhibits the protein-binding activity of the RNA Pol II C-terminal domain. PMID: 26515650
The amount of RNA polymerase II (RNAPII) on the HIV promoter and other viral regions was strongly diminished in HIV-infected CD4+ cells co-cultivated with cell non-cytotoxic antiviral response-expressing CD8+ cells. PMID: 26499373
Ash2L acts in concert with P53 promoter occupancy to activate RNA Polymerase II by aiding formation of a stable transcription pre-initiation complex required for its activation. PMID: 25023704
Data suggest that RNA polymerase II inhibitors may be a useful class of agent for targeting dormant leukaemia cells. PMID: 23767415
This viral pre-initiation complex is composed of five different proteins in addition to Epstein-Barr virus BcRF1 and interacts with cellular RNA polymerase II PMID: 25165108
Data show that E2F-1 form a complex with RNA polymerase II and protein PURA for transcriptional activation of the secondary promoter. PMID: 24819879
human CD68 gene expression is associated with changes in Pol II phosphorylation and short-range intrachromosomal gene looping PMID: 17583472
Authors show that the NSs protein of Schmallenberg virus (SBV) induces the degradation of the RPB1 subunit of RNA polymerase II and consequently inhibits global cellular protein synthesis and the antiviral response. PMID: 24828331
This study reveals that TCERG1 regulates HIV-1 transcriptional elongation by increasing the elongation rate of RNAPII and phosphorylation of Ser 2 within the carboxyl-terminal domain. PMID: 24165037
Slow Pol II elongation allows weak splice sites to be recognized, leading to higher inclusion of alternative exons. PMID: 24793692
sequence-specific double strand DNA breaks are sufficient to activate the positive transcription elongation factor b (P-TEFb), to trigger hyperphosphorylation of the largest RNA polymerase II carboxyl-terminal-domain (Rpb1-CTD) and to induce activation of p53-transcriptional axis resulting in cell cycle arrest. PMID: 23906511
interaction with nuclear CD26 and POLR2A gene PMID: 23638030
RECQL5 contacts the Rpb1 jaw domain of Pol II at a site that overlaps with the binding site for the transcription elongation factor TFIIS. Binding of RECQL5 to Pol II interferes with the ability of TFIIS to promote transcriptional read-through in vitro. PMID: 23748380
Data show that p68/DdX5 immunoprecipitated with RNA polymerase II (RNAP II) and suggest p68 is important in facilitating beta-catenin and androgen receptor (AR) transcriptional activity in prostate cancer cells. PMID: 23349811
inhibition of the transition of paused RNA PolII to productive elongation, described here for p21(CIP1), is a general mechanism by which transcription factor Sp3 fine-tunes gene expression. PMID: 23401853
RNA polymerase II acts as an RNA-dependent RNA polymerase to extend and destabilize a non-coding RNA. PMID: 23395899
Data indicate that polyamide treatment activates p53 signaling and results in a time- and and dose-dependent depletion of the RNA polymerase II (RNAP2) large subunit RPB1. PMID: 23319609
CTCF binding sites regulate mRNA production, RNA polymerase II (RNAPII) programming, and nucleosome organization of the Kaposi's sarcoma-associated herpesvirus latency transcript control region. PMID: 23192870
site-specific p65 phosphorylation targets NF-kappaB activity to particular gene subsets on a global level by influencing p65 and p-RNAP II promoter recruitment PMID: 23100252
BRD4-driven Pol II phosphorylation at serine 2 plays an important role in regulating lineage-specific gene transcription in human CD4+ T cells. PMID: 23086925
SNAPC1 is a general transcriptional coactivator that functions through elongating RNAPII. PMID: 22966203
Cyclin K1 is the primary cyclin partner for CDK12/CrkRS and it is required for activation of CDK12/CrkRS to phosphorylate the C-terminal domain of RNA Pol II. PMID: 22988298
Studies indicate that the super elongation complex (SEC) consisting of ELL, P-TEFb (CDK9) and MLL required for rapid transcriptional induction in the presence or absence of paused RNA polymerase II (Pol II). PMID: 22895430
Results indicate roles for both the RNA polymerase II C-terminal domain (CTD) and O-GlcNAc in the regulation of transcription initiation. PMID: 22605332
Here, the authors report phosphorylation of Thr4 by Polo-like kinase 3 in mammalian cells. PMID: 22549466
Studies suggest activator-induced structural shifts within Mediator trigger activation of stalled Pol II. PMID: 21326907
These results suggest that Mediator structural shifts induced by activator binding help stably orient pol II prior to transcription initiation within the human mediator-RNA polymerase II-TFIIF assembly. PMID: 22343046
evidence that phosphorylation of Rpb1 CTD Thr4 residues is required specifically for histone mRNA 3' end processing, functioning to facilitate recruitment of 3' processing factors to histone genes PMID: 22053051
Parcs/Gpn3 plays a critical role in the nuclear accumulation of RNAP II, and this function explains the relative importance of Parcs/Gpn3 in cell proliferation. PMID: 21782856
kinetics of RNA polymerase II elongation during co-transcriptional splicing PMID: 21264352
Data show that MicroRNA promoter identification based upon RPol II binding patterns provides important temporal and spatial measurements regarding the initiation of transcription. PMID: 21072189
The deregulation of cellular NIPP1/PP1 holoenzyme affects RNAPII phosphorylation and pointing to NIPP1 as a potential regulatory factor in RNAPII-mediated transcription. PMID: 20941529
Elevated PHD1 concomitant with decreased PHD2 are causatively related to Rpb1 hydroxylation and oncogenesis in human renal clear cell carcinomas with WT VHL gene. PMID: 20978146

Hide All

Subcellular Location

Nucleus. Cytoplasm. Chromosome.

Protein Families

RNA polymerase beta' chain family

Database Links

HGNC: 9187

OMIM: 180660

KEGG: hsa:5430

STRING: 9606.ENSP00000314949

UniGene: Hs.270017

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Code	CSB-RA018327A05phHU
Size	US$210
	Order now
Image	Western Blot Positive WB detected in Hela whole cell lysate,A549 whole cell lysate,293 whole cell lysate All lanes Phospho-POLR2A antibody at 0.75μg/ml Secondary Goat polyclonal to rabbit IgG at 1/50000 dilution Predicted band size: 270 KDa Observed band size: 270 KDa Immunocytochemistry analysis of CSB-RA018327A05phHU diluted at 1:75 and staining in Hela cells(treated with 100ng/ml EGF for 4h) performed on a Leica BondTM system. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system. Immunofluorescence staining of Hela cells with CSB-RA018327A05phHU at 1:100,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4℃. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).
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Phospho-POLR2A (S5) Recombinant Monoclonal Antibody

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Target Background