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Rat Recombinant Monoclonal RNA polymerase II RPB1 phospho S2 antibody. Suitable for ChIP, Dot, WB, I-ELISA, ICC/IF and reacts with Human, Mouse, Rat, Synthetic peptide - Human samples. Cited in 4 publications.

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Images

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (AB252855), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (AB252855), expandable thumbnail
  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (AB252855), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (AB252855), expandable thumbnail
  • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (AB252855), expandable thumbnail

Publications

Key facts

Isotype

IgG1

Host species

Rat

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIPDotWBI-ELISAICC/IF
Human
Tested
Expected
Tested
Expected
Tested
Mouse
Expected
Expected
Tested
Expected
Tested
Rat
Expected
Expected
Tested
Expected
Expected
Synthetic peptide - Human
Not recommended
Tested
Not recommended
Tested
Not recommended

Tested
Tested

Species

Human

Dilution info

5.00000-25.00000 µg chromatin

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide - Human

Dilution info

0.446 µg/mL

Notes

-

Expected
Expected

Species

Human, Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

0.446 µg/mL

Notes

-

Species

Rat

Dilution info

0.446 µg/mL

Notes

-

Species

Human

Dilution info

0.446 µg/mL

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide - Human

Dilution info

-

Notes

Use at 250 ng/ml.

Expected
Expected

Species

Human, Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

0.892 µg/mL

Notes

-

Species

Human

Dilution info

0.892 µg/mL

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

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

Rat Recombinant Monoclonal RNA polymerase II RPB1 phospho S2 antibody. Suitable for ChIP, Dot, WB, I-ELISA, ICC/IF and reacts with Human, Mouse, Rat, Synthetic peptide - Human samples. Cited in 4 publications.

Key facts

Isotype

IgG1

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

3E10

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RNA polymerase II CTD repeat YSPTSPS also known as the C-terminal domain of RNA polymerase II is a critical component of the RNA polymerase II enzyme commonly referred to as pol II. This domain is characterized by the repetitive sequence YSPTSPS which plays a significant role in the regulation of transcription. The mass of RNA polymerase II including its CTD varies but is essential for its function in gene expression. RNA polymerase II with the CTD is expressed in the nucleus of eukaryotic cells where it orchestrates the transcription of DNA into mRNA.

Biological function summary

RNA polymerase II CTD repeat YSPTSPS is essential for the transcription progression from initiation to termination. It is part of the large RNA polymerase II complex interacting with various transcription factors and enzymes necessary for RNA processing. The phosphorylation state of the CTD particularly on serine residues regulates interactions with splicing machinery and other RNA processing factors. This modulation ensures the coupling between transcription and RNA processing events controlling mRNA synthesis and maturation.

Pathways

RNA polymerase II CTD repeat YSPTSPS is important in the mRNA synthesis pathway specifically in transcriptional regulation and processing of nascent RNA transcripts. It interacts with proteins such as the transcription factors TFIIH and TFIIB which aid in promoter recognition and open complex formation. The CTD's dynamic phosphorylation pattern allows integration into multiple cellular pathways most importantly connecting transcription with RNA splicing and transport pathways.

Associated diseases and disorders

Abnormal function or mutations in RNA polymerase II CTD repeat YSPTSPS associate with diseases such as transcription-related syndromes and certain cancers. Deficient CTD phosphorylation can lead to improper mRNA processing resulting in neural developmental disorders. Additionally its interaction with proteins like CDK7 which phosphorylates the CTD links it to tumors where transcriptional dysregulation is a hallmark. Understanding the CTD's role in these diseases provides insight into therapeutic targets and strategies for intervention.

Product promise

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8 product images

  • ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Chromatin was prepared from HeLa cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
    The ChIP was performed with 25 μg of chromatin, 5 μg of ab252855 (red), or 5 μg of Rat IgG1 ab18407 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab252855 at 1/500 dilution, followed by Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line, the signal decreased after phosphatase treatment at 37°C for 2h. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 4 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310)

    Exposure time: 3 seconds

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855) at 1/1000 dilution

    Lane 1: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

    Lane 2: PC-12 whole cell lysate (Phosphatase treated membrane) at 10 µg

    Secondary

    All lanes: Goat Anti-Rat IgG (H+L), HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/5000 dilution

    Predicted band size: 217 kDa

    Observed band size: 260 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab252855 at 1/500 dilution, followed by Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in RAW 264.7 cell line, the signal decreased after phosphatase treatment at 37°C for 2h. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 4 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Dot blot analysis using ab252855 at 1/1000 dilution followed by a Goat Anti-Rat IgG (H+L), HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/5000 dilution. Blocking/diluting buffer and concentration: 5% NFDM/TBST. Exposure time: 3 minutes.

    Lane 1: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide
    Lane 2: RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide
    Lane 3: RNA polymerase II CTD repeat YSPTSPS (phospho T4) peptide
    Lane 4: RNA polymerase II CTD repeat YSPTSPS (phospho Y1) peptide
    Lane 5: RNA polymerase II CTD repeat YSPTSPS (phospho S7) peptide
    Lane 6: RNA polymerase II CTD repeat YSPTSPS non-phopho peptide

  • Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    ELISA analysis using ab252855 at a range of 1000-0 ng/ml followed by a Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rat IgG (H+L) at 1/1000 dilution. Antigen concentration: 1000 ng/ml

    Antigens: phospho S2, phospho S5, phospho T4, phospho Y1, phospho S7, non-phospho

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310)

    Exposure time: 7 seconds

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855) at 1/1000 dilution

    Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate (Untreated membrane) at 10 µg

    Lane 2: RAW264.7 whole cell lysate (Phosphatase treated membrane) at 10 µg

    Secondary

    All lanes: Goat Anti-Rat IgG (H+L), HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/5000 dilution

    Predicted band size: 217 kDa

    Observed band size: 260 kDa

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855), expandable thumbnail

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310)

    Exposure time: 3 seconds

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [3E10] (ab252855) at 1/1000 dilution

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (Untreated membrane) at 10 µg

    Lane 2: HeLa whole cell lysate (Phosphatase treated membrane) at 10 µg

    Secondary

    All lanes: Goat Anti-Rat IgG (H+L), HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/5000 dilution

    Predicted band size: 217 kDa

    Observed band size: 260 kDa

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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