Diagenode

Pol II Antibody

Catalog Number
Format
Price
C15200253-50
50 µg
$380.00
  Bulk order
Other format



Alternative names: POLR2A, RPB1, POLR2, RPOL2

Monoclonal antibody raised in mouse against the B1 subunit of RNA polymerase II (polymerase (RNA) II (DNA directed) polypeptide A) of wheat germ. Interacts with the highly conserved C-terminal domain of the protein containing the YSPTSPS repeat.

Lot001
Concentration1 µg/µl
Species reactivityHuman, Mouse, Xenopus: positive. Other species: not tested.
TypeMonoclonal. ChIP-grade. ChIP-seq grade.
PurityProtein A purified monoclonal antibody
HostMouse
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05 % Na-azide
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq* 0.5 - 1 µl per ChIP Fig 1, 2
Western Blotting 1:1,000 Fig 3

*Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 µg per IP.

  • Validation Data
    Pol II monoclonal antibody ChIP results

    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. C15200253) 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 0.5, 1, 2 and 5 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 of the GAPDH and EIF4A2 genes, used as positive controls, and for the MYOD1 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 2 Antibody ChIP-seq results

    Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II
    ChIP was performed on sheared chromatin from 1 million HeLa cells using 0.5 µg of the Diagenode antibody against Pol II (cat. No. C15200253) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1.5 Mb region of chromosome 1 (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes (fig 2C and D).

     

    Pol 2 antibody western blot results

    Figure 3. 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. C15200253) 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.

  •  Documents
    Pol II monoclonal antibody datasheet DATASHEET
    Monoclonal antibody raised in mouse against the B1 subunit of RNA polymerase II (polymerase (RNA)...
    Download
    Chromatin Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
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  •  Publications

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