PRODUCT CODE: ET1703-87

Recombinant Phospho POLR2A (S5) Monoclonal Antibody (ET1703-87)

  • Recombinant

Applications

  • WB

  • IP

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Phospho-POLR2A (S5) on different cells lysates using anti- Phospho-POLR2A (S5) antibody at 1/500 dilution.<br />
Positive control: <br />
 Line 1: Hela <br />
 Line 2: MCF-7
  • Western blot analysis of Phospho-POLR2A (S5) on different cells lysates using anti- Phospho-POLR2A (S5) antibody at 1/500 dilution.<br />
Positive control: <br />
 Line 1: Hela <br />
 Line 2: MCF-7
  • ICC staining Phospho-POLR2A (S5) in Hela cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • ICC staining Phospho-POLR2A (S5) in MCF-7 cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • ICC staining Phospho-POLR2A (S5) in PC-12 cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti- Phospho-POLR2A (S5) antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti- Phospho-POLR2A (S5) antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti- Phospho-POLR2A (S5) antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded mouse bladder tissue using anti- Phospho-POLR2A (S5) antibody. Counter stained with hematoxylin.
  • Flow cytometric analysis of Hela cells with Phospho-POLR2A (S5) antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Western blot analysis of Phospho-POLR2A (S5) on different cells lysates using anti- Phospho-POLR2A (S5) antibody at 1/500 dilution.
Positive control:
Line 1: Hela
Line 2: MCF-7

Applications

  • WB

  • IP

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho POLR2A (S5) Monoclonal Antibody (ET1703-87)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser5 of human polr2a.

Host

Rabbit

Modification

Phospho

Modification Site

S5

Positive Control

MCF-7, Hela, PC-12, human breast cancer tissue, human spleen tissue, human kidney tissue, mouse bladder tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JM51-21

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

250 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • ICC/IF

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

DNA-directed RNA polymerase II subunit RPB1

GENE NAME

POLR2A

SYNONYMS

RNA polymerase II subunit B1, POLR3A

SEQUENCE SIMILARITIES

Belongs to the RNA polymerase beta' chain family.

POST-TRANSLATIONAL MODIFICATION

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.; 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 (By similarity). Following UV treatment, the elongating form of RNA polymerase II (RNA pol IIo) is ubiquitinated on UV damage sites without leading to degradation: ubiquitination is facilitated by KIAA1530/UVSSA and promotes RNA pol IIo backtracking to allow access to the nucleotide excision repair machinery.

SUBCELLULAR LOCATION

Nucleus. Cytoplasm. Chromosome. Note=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.

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 (By similarity). 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 (By similarity).; (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.