Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18855-87] to RNA polymerase II CTD repeat YSPTSPS (phospho S2) - ChIP Grade
  • Suitable for: ChIP-sequencing, WB, Dot blot, IHC-P, ICC/IF, Flow Cyt, ChIP, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies

  • Description

    Rabbit monoclonal [EPR18855-87] to RNA polymerase II CTD repeat YSPTSPS (phospho S2) - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    ChIP-seq
    Human
    Flow Cyt
    Human
    ICC/IF
    Mouse
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, RAW 264.7 and PC-12 whole cell lysates. Dot blot: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide. IHC-P: Mouse testis tissue; rat testis tissue; human testis tissue. ICC/IF: HeLa and RAW 264.7 cells. Flow cyt: HeLa cells. IP: HeLa whole cell lysate. ChIP: Chromatin prepared from HeLa cells. ChIP-seq: Chromatin prepared from HeLa cells.

  • General 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 production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • ChIP-sequencing - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    ChIP-sequencing - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 4 µg of Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146). ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

    Additional screenshots of mapped reads can be downloaded here.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab238146 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat testis without alkaline phosphatase treatment (panel A). No signal was detected when treated with alkaline phosphatase (panel B). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab238146 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • ChIP-sequencing - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    ChIP-sequencing - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146). ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.

    Additional screenshots of mapped reads can be downloaded here.

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab238146 at 1/500 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary 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℃ for 2h. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Chromatin was prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10min.

    The ChIP was performed with 25 µg of chromatin, 5 µg of ab238146 (red), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).

    Primers and probes are located in the first kb of the transcribed region.

  • Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    RNA polymerase II CTD repeat YSPTSPS (phospho S2) was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab238146 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab238146 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

    Lane 1: HeLa whole cell lysate 10 μg (Input).
    Lane 2: ab237146 IP in HeLa whole cell lysate.
    Lane 3: : Rabbit monoclonal IgG (ab172730) instead of ab238146 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 7 seconds.

  • Flow Cytometry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Flow Cytometry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab238146 at 1/600 (red) compared with a Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146) at 1/5000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa whole cell lysate (phosphatase-treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 217 kDa
    Observed band size: 270 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes
  • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Dot blot analysis of RNA polymerase II CTD repeat YSPTSPS (phospho S2) labeled with ab238146 at 1/1000 dilution.

    Lane 1: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide.
    Lane 2: RNA polymerase II CTD repeat YSPTSPS non-phospho peptide.
    Lane 3: RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab238146 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis without alkaline phosphatase treatment (panel A). No signal was detected when treated with alkaline phosphatase (panel B). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab238146 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Immunohistochemical analysis of paraffin-embedded human testis tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab238146 at 1/2000 dilution, followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis without alkaline phosphatase treatment (panel A). No signal was detected when treated with alkaline phosphatase (panel B). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab238146 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab238146 at 1/500 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cell line, the signal decreased after phosphatase treatment at 37℃ for 2h. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146) at 1/2000 dilution

    Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 2 : RAW 264.7 whole cell lysate (phosphatase-treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 217 kDa
    Observed band size: 270 kDa why is the actual band size different from the predicted?


    Exposure time: 37 seconds


    Blocking and dilution buffer: 1% BSA/TBST.

    The molecular weight  observed is consistent with what has been described in the literature (PMID: 25849141).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146) at 1/2000 dilution

    Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 2 : PC-12 whole cell lysate (phosphatase-treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 217 kDa
    Observed band size: 270 kDa why is the actual band size different from the predicted?


    Exposure time: 37 seconds


    Blocking and dilution buffer: 1% BSA/TBST.

  • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)
    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (ab238146)

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