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Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide is a Synthetic blocking peptide. >90% purity and suitable for WB, BL.


Images

Dot Blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (AB18488), expandable thumbnail
  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (AB18488), expandable thumbnail
  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (AB18488), expandable thumbnail
  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (AB18488), expandable thumbnail
  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (AB18488), expandable thumbnail

Publications

Key facts

Purity

>90% HPLC

Tags

Tag free

Applications

WB, BL

Biologically active

No

Reactivity data

Application

WB

Reactivity

Reacts

Dilution info

-

Notes

-

Application

BL

Reactivity

Reacts

Dilution info

-

Notes

This peptide can be used with studies using Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody ab5131.

Associated Products

Select an associated product type

2 products for Alternative Product

Target data

Function

Catalytic core component of RNA polymerase II (Pol II), a DNA-dependent RNA polymerase which synthesizes mRNA precursors and many functional non-coding RNAs using the four ribonucleoside triphosphates as substrates (By similarity) (PubMed:23748380, PubMed:27193682, PubMed:30190596, PubMed:9852112). Pol II-mediated transcription cycle proceeds through transcription initiation, transcription elongation and transcription termination stages. During transcription initiation, Pol II pre-initiation complex (PIC) is recruited to DNA promoters, with focused-type promoters containing either the initiator (Inr) element, or the TATA-box found in cell-type specific genes and dispersed-type promoters that often contain hypomethylated CpG islands usually found in housekeeping genes. Once the polymerase has escaped from the promoter it enters the elongation phase during which RNA is actively polymerized, based on complementarity with the template DNA strand. Transcription termination involves the release of the RNA transcript and polymerase from the DNA (By similarity) (PubMed:23748380, PubMed:27193682, PubMed:28108474, PubMed:30190596, PubMed:9852112). Forms Pol II active center together with the second largest subunit POLR2B/RPB2. Appends one nucleotide at a time to the 3' end of the nascent RNA, with POLR2A/RPB1 most likely contributing a Mg(2+)-coordinating DxDGD motif, and POLR2B/RPB2 participating in the coordination of a second Mg(2+) ion and providing lysine residues believed to facilitate Watson-Crick base pairing between the incoming nucleotide and template base. Typically, Mg(2+) ions direct a 5' nucleoside triphosphate to form a phosphodiester bond with the 3' hydroxyl of the preceding nucleotide of the nascent RNA, with the elimination of pyrophosphate. The reversible pyrophosphorolysis can occur at high pyrophosphate concentrations (By similarity) (PubMed:30190596, PubMed:8381534, PubMed:9852112). Can proofread the nascent RNA transcript by means of a 3' -> 5' exonuclease activity. If a ribonucleotide is mis-incorporated, backtracks along the template DNA and cleaves the phosphodiester bond releasing the mis-incorporated 5'-ribonucleotide (By similarity) (PubMed:8381534). Through its unique C-terminal domain (CTD, 52 heptapeptide tandem repeats) serves as a platform for assembly of factors that regulate transcription initiation, elongation and termination. CTD phosphorylation on Ser-5 mediates Pol II promoter escape, whereas phosphorylation on Ser-2 is required for Pol II pause release during transcription elongation and further pre-mRNA processing. Additionally, the regulation of gene expression levels depends on the balance between methylation and acetylation levels of the CTD-lysines. Initiation or early elongation steps of transcription of growth-factor-induced immediate early genes are regulated by the acetylation status of the CTD. Methylation and dimethylation have a repressive effect on target genes expression. Cooperates with mRNA splicing machinery in co-transcriptional 5'-end capping and co-transcriptional splicing of pre-mRNA (By similarity) (PubMed:24207025, PubMed:26124092).RNA-dependent RNA polymerase that catalyzes the extension of a non-coding RNA (ncRNA) at the 3'-end using the four ribonucleoside triphosphates as substrates. An internal ncRNA sequence near the 3'-end serves as a template in a single-round Pol II-mediated RNA polymerization reaction. May decrease the stability of ncRNAs that repress Pol II-mediated gene transcription.(Microbial infection) Acts as an RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicase and transcriptase for the viral RNA circular genome.

Alternative names

Recommended products

Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide is a Synthetic blocking peptide. >90% purity and suitable for WB, BL.

Key facts

Purity

>90% HPLC

Applications

WB, BL

Accession
P24928-1
Animal free

No

Species

Human

Concentration
Loading...
Storage buffer

Constituents: HEPES, 0.87% Sodium chloride, 0.0584% EDTA, 0.001% Sorbitan monolaurate, ethoxylated

Sequence info

Amino acid sequence

Accession

P24928

Nature

Synthetic

Specifications

Form

Liquid

General info

Function

Catalytic core component of RNA polymerase II (Pol II), a DNA-dependent RNA polymerase which synthesizes mRNA precursors and many functional non-coding RNAs using the four ribonucleoside triphosphates as substrates (By similarity) (PubMed:23748380, PubMed:27193682, PubMed:30190596, PubMed:9852112). Pol II-mediated transcription cycle proceeds through transcription initiation, transcription elongation and transcription termination stages. During transcription initiation, Pol II pre-initiation complex (PIC) is recruited to DNA promoters, with focused-type promoters containing either the initiator (Inr) element, or the TATA-box found in cell-type specific genes and dispersed-type promoters that often contain hypomethylated CpG islands usually found in housekeeping genes. Once the polymerase has escaped from the promoter it enters the elongation phase during which RNA is actively polymerized, based on complementarity with the template DNA strand. Transcription termination involves the release of the RNA transcript and polymerase from the DNA (By similarity) (PubMed:23748380, PubMed:27193682, PubMed:28108474, PubMed:30190596, PubMed:9852112). Forms Pol II active center together with the second largest subunit POLR2B/RPB2. Appends one nucleotide at a time to the 3' end of the nascent RNA, with POLR2A/RPB1 most likely contributing a Mg(2+)-coordinating DxDGD motif, and POLR2B/RPB2 participating in the coordination of a second Mg(2+) ion and providing lysine residues believed to facilitate Watson-Crick base pairing between the incoming nucleotide and template base. Typically, Mg(2+) ions direct a 5' nucleoside triphosphate to form a phosphodiester bond with the 3' hydroxyl of the preceding nucleotide of the nascent RNA, with the elimination of pyrophosphate. The reversible pyrophosphorolysis can occur at high pyrophosphate concentrations (By similarity) (PubMed:30190596, PubMed:8381534, PubMed:9852112). Can proofread the nascent RNA transcript by means of a 3' -> 5' exonuclease activity. If a ribonucleotide is mis-incorporated, backtracks along the template DNA and cleaves the phosphodiester bond releasing the mis-incorporated 5'-ribonucleotide (By similarity) (PubMed:8381534). Through its unique C-terminal domain (CTD, 52 heptapeptide tandem repeats) serves as a platform for assembly of factors that regulate transcription initiation, elongation and termination. CTD phosphorylation on Ser-5 mediates Pol II promoter escape, whereas phosphorylation on Ser-2 is required for Pol II pause release during transcription elongation and further pre-mRNA processing. Additionally, the regulation of gene expression levels depends on the balance between methylation and acetylation levels of the CTD-lysines. Initiation or early elongation steps of transcription of growth-factor-induced immediate early genes are regulated by the acetylation status of the CTD. Methylation and dimethylation have a repressive effect on target genes expression. Cooperates with mRNA splicing machinery in co-transcriptional 5'-end capping and co-transcriptional splicing of pre-mRNA (By similarity) (PubMed:24207025, PubMed:26124092).

Sequence similarities

Belongs to the RNA polymerase beta' chain family.

Post-translational modifications

The tandem heptapeptide repeats in the C-terminal domain (CTD) can be highly phosphorylated (PubMed:28076779). 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.

Subcellular localisation

Nucleus

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

This peptide is 2 repeats of the YSPTSPS repeated motif found at the C-terminal of RNA polymerase II.

- First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
- If the peptide doesn't dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
- Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
- Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
- Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.

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5 product images

  • Dot Blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488), expandable thumbnail
    This image is courtesy of an anonymous customer review

    Dot Blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

    Dot blot analysis of ab18488 - RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5.

    0.1 μg peptide was spotted onto the membrane and blocked with 2% BSA for 1 hour at room temperature, before detection with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody ab5131 at 1/3000 dilution.

  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488), expandable thumbnail

    Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody ab5131) at 1 µg/mL

    Lanes 1, 3, 5 and 7: HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg

    Lanes 2, 4, 6 and 8: S.cerevisiae (Y190) Whole Cell Lysate at 20 µg

    Secondary

    All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 217 kDa

    Exposure time: 30s

  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488), expandable thumbnail

    Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

    Lanes 1, 3 and 4: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody ab5131) at 1/500 dilution

    Lane 2: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody ab5131) at 1/2000 dilution

    All lanes: HeLa nuclear extract at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 217 kDa

    Observed band size: 250 kDa

    Exposure time: 30s

  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488), expandable thumbnail

    Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody ab5095) at 1 µg/mL

    Lanes 1, 3, 5 and 7: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Lanes 2, 4, 6 and 8: S.cerevisiae (Y190) Whole Cell Lysate at 10 µg

    Secondary

    All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 217 kDa

    Observed band size: 240 kDa

  • Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488), expandable thumbnail

    Western blot - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488)

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (Anti-RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade ab26721) at 1 µg/mL

    Lanes 1, 3, 5 and 7: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

    Lanes 2, 4, 6 and 8: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

    Secondary

    All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 217 kDa

    Observed band size: 220 kDa

    Exposure time: 1min

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Product protocols

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