• Immunoprecipitation analysis of 200ug extracts of HeLa cells using 3ug POLR2A Rabbit polyclonal antibody (STJ25045 1:100). Western blot was performed from the immunoprecipitate using POLR2A Rabbit polyclonal antibody (STJ25045) at a dilition of 1:1000.
  • Western blot analysis of various lysates, using POLR2A Rabbit polyclonal antibody (STJ25045) at 1:2000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Negative control (NC) : Human Plasma. Exposure time: 30s.

Anti-POLR2A antibody (1-1970) (STJ25045)

SKU:
STJ25045

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Host: Rabbit
Applications: WB/IF/IP
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-POLR2A (1-1970) is suitable for use in Western Blot, Immunofluorescence and Immunoprecipitation research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.01% Thimerosal, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:500-1:2000
IF/ICC 1:50-1:200
IP 1:50-1:200
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: POLR2A
Gene ID: 5430
Uniprot ID: RPB1_HUMAN
Immunogen Region: 1-1970
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 1-1970 of human POLR2A (NP_000928.1).
Immunogen Sequence: MHGGGPPSGDSACPLRTIKR VQFGVLSPDELKRMSVTEGG IKYPETTEGGRPKLGGLMDP RQGVIERTGRCQTCAGNMTE CPGHFGHIELAKPVFHVGFL VKTMKVLRCVCFFCSKLLVD SNNPKIKDILAKSKGQPKKR LTHVYDLCKGKNICEGGEEM DNKFGVEQPEGDEDLTKEKG HGGCGRYQPRIRRSGLELYA EWKHVNEDSQEKKILLSPER VHEIFKRISDEECFVLGME
Post Translational Modifications The tandem heptapeptide 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 heptapeptide repeat and is mediated, at least, by CDK7 and CDK9. CDK7 phosphorylation of POLR2A associated with DNA promotes transcription initiation by triggering dissociation from DNA. Phosphorylation also takes place at 'Ser-7' of the heptapeptide repeat, which is required for efficient transcription of snRNA genes and processing of the transcripts. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphatases, and a 'CTD code' that specifies the position of Pol II within the transcription cycle has been proposed. Dephosphorylated by the protein phosphatase CTDSP1. Dephosphorylated at 'Ser-2' following UV irradiation. Among tandem heptapeptide repeats of the C-terminal domain (CTD) some do not match the Y-S-P-T-S-P-S consensus, the seventh serine residue 'Ser-7' being replaced by a lysine. 'Lys-7' in these non-consensus heptapeptide repeats can be alternatively acetylated, methylated and dimethylated. EP300 is one of the enzyme able to acetylate 'Lys-7'. Acetylation at 'Lys-7' of non-consensus heptapeptide repeats is associated with 'Ser-2' phosphorylation and active transcription. Regulates initiation or early elongation steps of transcription specially for inducible genes. Methylated at Arg-1810 prior to transcription initiation when the CTD is hypophosphorylated, phosphorylation at Ser-1805 and Ser-1808 preventing this methylation. Symmetrically or asymmetrically dimethylated at Arg-1810 by PRMT5 and CARM1 respectively. Symmetric or asymmetric dimethylation modulates interactions with CTD-binding proteins like SMN1/SMN2 and TDRD3. SMN1/SMN2 interacts preferentially with the symmetrically dimethylated form while TDRD3 interacts with the asymmetric form. Through the recruitment of SMN1/SMN2, symmetric dimethylation is required for resolving RNA-DNA hybrids created by RNA polymerase II, that form R-loop in transcription terminal regions, an important step in proper transcription termination. CTD dimethylation may also facilitate the expression of select RNAs. Among tandem heptapeptide repeats of the C-terminal domain (CTD) some do not match the Y-S-P-T-S-P-S consensus, the seventh serine residue 'Ser-7' being replaced by a lysine. 'Lys-7' in these non-consensus heptapeptide repeats can be alternatively acetylated, methylated, dimethylated and trimethylated. Methylation occurs in the earliest transcription stages and precedes or is concomitant to 'Ser-5' and 'Ser-7' phosphorylation. Dimethylation and trimehtylation at 'Lys-7' of non-consensus heptapeptide repeats are exclusively associated with phosphorylated CTD. Ubiquitinated by WWP2 leading to proteasomal degradation. Following transcription stress, the elongating form of RNA polymerase II (RNA pol IIo) is ubiquitinated by NEDD4 on Lys-1268 at DNA damage sites without leading to degradation: ubiquitination promotes RNA pol IIo backtracking to allow access by the transcription-coupled nucleotide excision repair (TC-NER) machinery. At stalled RNA pol II where TC-NER has failed, RBX1-mediated polybiquitination at Lys-1268 may lead to proteasome-mediated degradation in a UBAP2- and UBAP2L-dependent manner.presumably to halt global transcription and enable 'last resort' DNA repair pathways.
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.
Protein Name Dna-Directed Rna Polymerase Ii Subunit Rpb1
Rna Polymerase Ii Subunit B1
Dna-Directed Rna Polymerase Ii Subunit A
Dna-Directed Rna Polymerase Iii Largest Subunit
Rna-Directed Rna Polymerase Ii Subunit Rpb1
Database Links Reactome: R-HSA-112382
Reactome: R-HSA-113418
Reactome: R-HSA-167152
Reactome: R-HSA-167158
Reactome: R-HSA-167160
Reactome: R-HSA-167161
Reactome: R-HSA-167162
Reactome: R-HSA-167172
Reactome: R-HSA-167200
Reactome: R-HSA-167238
Reactome: R-HSA-167242
Reactome: R-HSA-167243
Reactome: R-HSA-167246
Reactome: R-HSA-167287
Reactome: R-HSA-167290
Reactome: R-HSA-168325
Reactome: R-HSA-203927
Reactome: R-HSA-5578749
Reactome: R-HSA-5601884
Reactome: R-HSA-5617472
Reactome: R-HSA-674695
Reactome: R-HSA-6781823
Reactome: R-HSA-6781827
Reactome: R-HSA-6782135
Reactome: R-HSA-6782210
Reactome: R-HSA-6796648
Reactome: R-HSA-6803529
Reactome: R-HSA-6807505
Reactome: R-HSA-72086
Reactome: R-HSA-72163
Reactome: R-HSA-72165
Reactome: R-HSA-72203
Reactome: R-HSA-73776
Reactome: R-HSA-73779
Reactome: R-HSA-75953
Reactome: R-HSA-75955
Reactome: R-HSA-76042
Reactome: R-HSA-77075
Reactome: R-HSA-8851708
Reactome: R-HSA-9018519
Reactome: R-HSA-9670095
Cellular Localisation Nucleus
Cytoplasm
Chromosome
Hypophosphorylated Form Is Mainly Found In The Cytoplasm
While The Hyperphosphorylated And Active Form Is Nuclear
Co-Localizes With Kinase Srpk2 And Helicase Ddx23 At Chromatin Loci Where Unscheduled R-Loops Form
Alternative Antibody Names Anti-Dna-Directed Rna Polymerase Ii Subunit Rpb1 antibody
Anti-Rna Polymerase Ii Subunit B1 antibody
Anti-Dna-Directed Rna Polymerase Ii Subunit A antibody
Anti-Dna-Directed Rna Polymerase Iii Largest Subunit antibody
Anti-Rna-Directed Rna Polymerase Ii Subunit Rpb1 antibody
Anti-POLR2A antibody
Anti-POLR2 antibody

Information sourced from Uniprot.org

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