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Mouse Monoclonal RNA polymerase II RPB1 phospho S5 antibody. Suitable for ChIP, Flow Cyt (Intra), ELISA, Dot, WB, ICC/IF and reacts with Human, Mouse, Rat, Synthetic peptide - Human samples. Cited in 362 publications.


Images

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (AB5408), expandable thumbnail
  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (AB5408), expandable thumbnail
  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (AB5408), expandable thumbnail
  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (AB5408), expandable thumbnail
  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (AB5408), expandable thumbnail

Publications

  • Nature communications 14:80492023
    RIF1 regulates early replication timing in murine B cells.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Daniel Malzl,Mihaela Peycheva,Ali Rahjouei,Stefano Gnan,Kyle N Klein,Mariia Nazarova,Ursula E Schoeberl,David M Gilbert,Sara C B Buonomo,Michela Di Virgilio,Tobias Neumann,Rushad Pavri
    PubMed 38081811
  • PLoS genetics 19:e10110212023
    Deletions of singular U1 snRNA gene significantly interfere with transcription and 3'-end mRNA formation.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Mei Wang,An-Min Liang,Zhen-Zhen Zhou,Ting-Lin Pang,Yu-Jie Fan,Yong-Zhen Xu
    PubMed 37917726

Key facts

Isotype

IgG1

Host species

Mouse

Storage buffer

pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIPFlow Cyt (Intra)ELISADotWBICC/IF
Human
Tested
Tested
Expected
Expected
Tested
Tested
Mouse
Expected
Expected
Expected
Expected
Tested
Expected
Rat
Expected
Expected
Expected
Expected
Tested
Expected
Arabidopsis thaliana
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Synthetic peptide - Human
Not recommended
Not recommended
Tested
Tested
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1-4 µg for 106 Cells

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Arabidopsis thaliana

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1 µg for 106 Cells

Notes

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Arabidopsis thaliana

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Expected
Expected

Species

Human, Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Arabidopsis thaliana

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide - Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Human, Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Arabidopsis thaliana

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Predicted
Predicted

Species

Arabidopsis thaliana

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Arabidopsis thaliana

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 products for Alternative Version

7 products for Alternative Product

Target data

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 (PubMed:24207025). 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.

Alternative names

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Mouse Monoclonal RNA polymerase II RPB1 phospho S5 antibody. Suitable for ChIP, Flow Cyt (Intra), ELISA, Dot, WB, ICC/IF and reacts with Human, Mouse, Rat, Synthetic peptide - Human samples. Cited in 362 publications.

Alternative names

Key facts

Isotype

IgG1

Form

Liquid

Clonality

Monoclonal

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

4H8

Purification technique

Affinity purification Protein G

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

ChIP protocols:
ChIP protocol for cross-linking ChIP (X-ChIP)
Native ChIP protocol
Chromatin preparation from tissues for ChIP
ChIP troubleshooting
ChIP tips and tricks guide

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

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

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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9 product images

  • ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    Chromatin was prepared from U-2 OS (human bone osteosarcoma epithelial cell line) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 μg of chromatin, 2 μg of ab5408 (blue), and 20 μl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    Blocking/Diluting Buffer and concentration

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408) at 1/1000 dilution

    Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 10 µg

    Lane 2: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate. Then the membrane was incubated with alkaline phosphatase at 10 µg

    Secondary

    All lanes: HRP-conjugated goat anti-mouse IgG at 1/10000 dilution

    Predicted band size: 217 kDa

    Observed band size: 270 kDa

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408) at 1/1000 dilution

    Lane 1: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate (ab28419) at 10 µg

    Lane 2: NIH 3T3 (Mouse) Whole Cell Lysate (ab52956) at 10 µg

    Lane 3: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957) at 10 µg

    Lane 4: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate (ab28419) Then the membrane was incubated with alkaline phosphatase at 10 µg

    Lane 5: NIH 3T3 (Mouse) Whole Cell Lysate (ab52956) Then the membrane was incubated with alkaline phosphatase at 10 µg

    Lane 6: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957) Then the membrane was incubated with alkaline phosphatase at 10 µg

    Secondary

    All lanes: HRP-conjugated goat anti-mouse IgG at 1/10000 dilution

    Predicted band size: 217 kDa

    Observed band size: 270 kDa

    Exposure time: 8min

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    Blocking/Diluting Buffer and concentration 5% NFDM/TBST

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408) at 1/1000 dilution

    Lane 1: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

    Lane 2: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate. Then the membrane was incubated with alkaline phosphatase at 10 µg

    Secondary

    All lanes: HRP-conjugated goat anti-mouse IgG at 1/10000 dilution

    Predicted band size: 217 kDa

    Observed band size: 270 kDa

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail
    This image is courtesy of an anonymous customer review.

    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    7.5% SDS gel.

    Blocked with 5% BSA for 1 hour at 25°C.

    Incubated with the primary antibody for 3 hours at 25°C in 1X TBST buffer.

    All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408) at 1/800 dilution

    Lane 1: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg

    Lane 2: HEK293T transfected with Tat containing plasmid whole cell lysate at 30 µg

    Secondary

    All lanes: HRP-conjugated goat anti-mouse IgG at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 217 kDa

    Observed band size: 260 kDa

    Exposure time: 30s

  • ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    ELISA using ab5408 at varying antibody concentrations.

    Curve_SPL4 indicates binding to RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5 (Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide ab18488).

    Binding to the following peptides was much weaker: Curve_SPL5 RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2 (S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide ab12793), Curve_SPL6 RNA polymerase II CTD repeat YSPTSPS peptide (ab12795).

  • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    Dot blot analysis of RNA polymerase II CTD repeat YSPTSPS (phospho S5) phospho peptide (Lane 1), RNA polymerase II CTD repeat YSPTSPS (phospho 2) phospho peptide (Lane 2) and RNA polymerase II CTD repeat YSPTSPS non-phospho peptide (Lane 3) labeling RNA polymerase II CTD repeat YSPTSPS (phospho S5) phospho peptide with ab5408 at a dilution of 1/1000 dilution (1ug/ml). A HRP-conjugated goat anti-mouse IgG was used as the secondary antibody at a dilution of 1/10,000 dilution.

    Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST .

  • Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab5408 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5408, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408)

    ab5408 staining RNA polymerase II CTD repeat YSPTSPS (phospho S5) in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab5408 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

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Product protocols

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