Anti-RPB1 | RNA polymerase II subunit B1

Product no: AS11 1804

AS11 1804 Clonality: Polyclonal Host: Rabbit | Reactivity:Arabidopsis thaliana

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  • Product Info
  • Immunogen:

    KLH-conjugated synthetic peptide derived from known higher plant polymerase II sequences, including Arabidopsis thaliana P18616, At4g35800

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl of sterile water
    Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Chromatin Immuno Precipitation (ChIP), Immunoprecipitation (IP)
    Recommended dilution: 1,2 or 5 µg (ChIP, IP)
    Expected | apparent MW:

    204 kDa

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Amygdalus persica, Asparagus officinalis, Brassica rapa, Capsella rubella, Glycine max, Hordeum vulgare, Solanum lycopersicum, Oryza glaberrima, Panicum italicum, Persea americana, Pisum sativum, Ricinus communis, Solanum tuberosum, Sorghum vulgare, Vitis vinifera
    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example

    IP using anti-plant polymerase II antibodies

    Chromatin immunoprecipitation (ChIP). Chromatin immunoprecipitation was performed as described (Bowler et al. (2004), Plant Journal) with modifications. Sonic and IP buffer were prepared as described (Kaufmann et al. (2010)), Nature Protocols). Chromatin pellet was sonicated at 4 °C with a Diagenode Bioruptor set at high intensity for 10 min (30 sec on, 30 sec off intervals) to obtain 250-500 bp DNA fragments. Chromatin (20 µg) was immunoprecipitated with a RNA Pol II antibody (2µg/IP)and Dynabeads®Protein G (Life Technologies). Reverse corsslinking and elution of DNA fragments were performed using Chelex (Biorad) at 95°C, 10 min as described (Rowley et al. (2013), Methods). Sample without antibody was used to determine the nonspecific background pulled-down directly by the beads. Relative abundance of regions of interest in immunoprecipitated DNA was measured by real-time PCR. Red lines indicates amplified region. Values on the charts are shown as the mean ± SD level (% of input) of RNA Pol II on GAPDH gene (A) and MIR168a (B) gene. Data comes from three independent experiments.

    Courtesy of MSc Jakub Dolata, Adam Mickiewicz University, Poland

  • Additional Information
  • Additional information: Purified IgG was lyophilized from PBS pH 7,4 + 0,02 % sodium azide, Therefore only distilled water is required for its re-constitution
    Additional information (application): This antibody is not suitable for western blot in denatured conditions
  • Background
  • Background:

    DNA-directed RNA polymerase II subunit 1 (EC= catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. This enzyme is the central component of the basal RNA polymerase II transcription machinery. Synonymes:DNA-directed RNA polymerase III largest subunit,DNA-directed RNA polymerase II subunit RPB1.

  • Product Citations
  • Selected references: Crawford et al. (2024).The Mediator kinase module enhances polymerase activity to regulate transcriptional memory after heat stress in Arabidopsis. EMBO J. 2024 Feb;43(3):437-461.  
    Wang et al. (2020). Close arrangement of CARK3 and PMEIL affects ABA-mediated pollen sterility in Arabidopsis thaliana. Plant Cell Environ. 2020 Aug 20.doi: 10.1111/pce.13871.
    Godoy Herz et al. (2019). Light Regulates Plant Alternative Splicing through the Control of Transcriptional Elongation. Mol Cell. 2019 Jan 15. pii: S1097-2765(18)31035-9. doi: 10.1016/j.molcel.2018.12.005.
    Dolata et al. (2015). NTR1 is required for transcription elongation checkpoints at alternative exons in Arabidopsis. EMBO J. 2015 Jan 7. pii: e201489478.
  • Protocols
  • Protocols to work with plant and algal protein extracts

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