Diagenode

Pol II monoclonal antibody - Classic (sample size)

Histone-Deacetylase-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15200004-10
10 µl
$80.00
  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. 

Lot001-11
Concentration1.0 µg/µl
Species reactivityHuman, Xenopus
TypeMonoclonal
PurityProtein A purified
HostMouse
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1 μg/IP Fig 1, 2
ELISA 1:3,000 Fig 3
Western Blotting 1:1,000 Fig 4
Immunofluorescence 1:500 Fig 5

* 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

    ChIP

    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. AB-001-0024), 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).

    ChIP-seq figure A

    ChIP-seq figure B

    ChIP-seq figure C

    ChIP-seq figure D

    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).

    ELISA

    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.

    Western blot

    Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II
    Nuclear 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.

    Immunofluorescence

    Figure 5. 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.

  • Applications
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    WB
    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
    IF
    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)
    Read more
  • Documents
    Datasheet Polll C15200004 DATASHEET
    Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymeras...
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    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Pol II monoclonal antibody - Classic (sample size) (Diagenode Cat# C15200004-10 Lot# 001-11). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Functional incompatibility between the generic NF-κB motif and a subtype-specific Sp1III element drives the formation of HIV-1 subtype C viral promoter
    Verma A et al.
    Of the various genetic subtypes of HIV-1, HIV-2 and SIV, only in subtype C of HIV-1, a genetically variant NF-κB binding site is found at the core of the viral promoter in association with a subtype-specific Sp1III motif. How the subtype-associated variations in the core transcription factor binding sites (TFB...

    Embryonic transcription is controlled by maternally defined chromatin state
    Hontelez S et al.
    Histone-modifying enzymes are required for cell identity and lineage commitment, however little is known about the regulatory origins of the epigenome during embryonic development. Here we generate a comprehensive set of epigenome reference maps, which we use to determine the extent to which maternal factors shape c...

    iRNA-seq: computational method for genome-wide assessment of acute transcriptional regulation from total RNA-seq data.
    Madsen JG, Schmidt SF, Larsen BD, Loft A, Nielsen R, Mandrup S
    RNA-seq is a sensitive and accurate technique to compare steady-state levels of RNA between different cellular states. However, as it does not provide an account of transcriptional activity per se, other technologies are needed to more precisely determine acute transcriptional responses. Here, we have developed an e...

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