Pol II monoclonal antibody

Catalog Number
50 μg
  Bulk order
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Alternative names: POLR2A, RPB1, POLR2, RPOL2

Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II. 

Concentration1.0 µg/µl
Species reactivityHuman, Xenopus, Yeast: positive. Other species: not tested.
TypeMonoclonal ChIP grade, ChIP-seq grade
PurityProtein A purified monoclonal antibody.
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05% azide.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 1 µg/ChIP Fig 1, 2
ELISA 1:3,000 Fig 3
Western Blotting 1:1,000 Fig 4, 5
Immunofluorescence 1:500 Fig 6

* Please note that the optimal antibody amount per ChIP should be determined by the end-user. We recommend testing 1-5 µg per IP.

  • Validation Data
    Pol II Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II
    ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II (Cat. No. C15200004) and optimized PCR primer pairs for qPCR. ChIP was performed with the "iDeal ChIP-seq" kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (2 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    Pol II Antibody ChIP-seq Grade
    Pol II Antibody for ChIP-seq
    Pol II Antibody for ChIP-seq assay
    Pol II Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II
    ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of the Diagenode antibody against Pol II (Cat. No. C15200004) as described above. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 400 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).

    Pol II Antibody ELISA validation

    Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol II
    To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol II (Cat. No. C15200004). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows that the antibody recognizes the unphosphorylated Pol II as well as most phosphorylated forms.

    Pol II Antibody for Western Blot

    Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol IINuclear extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II (Cat. No. C15200004) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

    Pol II Antibody validated in Western Blot

    Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II
    Whole cell extracts (40 µg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II (Cat. No. C15200004) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

    Pol II Antibody for Immunofluorescence

    Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II
    HeLa cells were stained with the Diagenode antibody against Pol II (Cat. No. C15200004) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • Target Description

    RNA polymerase II (pol II) is a key enzyme in the regulation and control of gene transcription. It is able to unwind the DNA double helix, synthesize RNA, and proofread the result. Pol II is a complex enzyme, consisting of 12 subunits, of which the B1 subunit (UniProt/Swiss-Prot entry P24928) is the largest. Together with the second largest subunit, B1 forms the catalytic core of the RNA polymerase II transcription machinery.

  •  お客様の声

    I have used  Diagenode's antibodies for my ChIP Seq experiments. The antibodies performed very well in our experiments with specific signal, and good signal to noise ratio.

    Antibodies used in our lab: Pol II Monoclonal Classic (C15200004).

    Junaid Akhtar, Institute of Development Biology and Neurobiology, Mainz, Germany
  •  実験手法
    Enzyme-linked immunosorbent assay. Read more
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
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    siRNA Knockdown
    Epigenetic antibodies you can trust! Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level o... Read more
  •  資料
    Datasheet Polll C15200004 DATASHEET
    Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymeras...
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
  •  Safety sheets
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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Pol II monoclonal antibody (Diagenode Cat# C15200004 Lot# 001-14). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

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