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Pol II S5p monoclonal antibody

Purified Mouse Monoclonal Antibody

     
  • CHIP - Pol II S5p monoclonal antibody ADN10008
    ChIP results obtained with the monoclonal antibody directed against Pol II S5p
    ChIP assays were performed using human HeLa cells, the monoclonal antibody against Pol II S5p and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit, 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).
  • E - Pol II S5p monoclonal antibody ADN10008
    Cross reactivity of the monoclonal antibody directed against Pol IIS5p
    To test the specificity an ELISA was performed using a serial dilution of the monoclonal antibody against Pol IIS5p. The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.
  • WB - Pol II S5p monoclonal antibody ADN10008
    Western blot analysis using the monoclonal antibody directed against Pol II S5p
    Nuclear extracts (25 µg) from HeLa cells were analysed by Western blot using the monoclonal antibody against Pol II S5p 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.
  • IF - Pol II S5p monoclonal antibody ADN10008
    Immunofluorescence using the monoclonal antibody directed against Pol II S5p
    HeLa cells were stained with the antibody against Pol II S5p (Cat. No. C15200007) 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 S5p 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.
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IF, E
Primary Accession P24928
Reactivity Human
Host Mouse
Clonality Monoclonal
Calculated MW 217176 Da
Additional Information
Gene ID 5430
Other Names DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II subunit B1, 2.7.7.6, DNA-directed RNA polymerase II subunit A, DNA-directed RNA polymerase III largest subunit, RNA-directed RNA polymerase II subunit RPB1, 2.7.7.48, POLR2A, POLR2
Target/Specificity Pol II S5p
PrecautionsPol II S5p monoclonal antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name POLR2A
Function DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single-stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as an RNA- dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.
Cellular Location Nucleus.
Citations (0)

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Background

DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single-stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as an RNA- dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.

References

Wintzerith M.,et al.Nucleic Acids Res. 20:910-910(1992).
Mita K.,et al.Gene 159:285-286(1995).
Zody M.C.,et al.Nature 440:1045-1049(2006).
Mural R.J.,et al.Submitted (SEP-2005) to the EMBL/GenBank/DDBJ databases.
Kershnar E.,et al.J. Biol. Chem. 273:34444-34453(1998).

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$ 290.00
Cat# ADN10008
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