POLR2A Antibody (OADC00206)

• Product Number: OADC00206
• Name: POLR2A Antibody (OADC00206)
• Isotype: IgG1
• Conjugation: Unconjugated
• NCBI Gene Id: 5430;
• Host: Mouse
• Clonality: Monoclonal
• Concentration: 1 mg/mL
• Gene Full Name: polymerase (RNA) II subunit A
• Alias Symbols: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, NEDHIB, RpIILS, hsRPB1, hRPB220
• Product Format: Liquid. Provided in PBS containing 0.05% azide.
• Description of Target: This gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. The product of this gene contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, this subunit, in combination with several other polymerase subunits, forms the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA.
• Reconstitution and Storage: Store at -20C. Avoid repeated freeze/thaw cycles.
• Specificity: Mouse monoclonal antibody raised against POLR2A
• Application Info: CHIP: 1-2 ug/CHIP
  ELISA: 1:3,000
  Western Blotting: 1:1,000
  Immunofluorescence: 1:500
• Uniprot ID: P24928
• Protein Name: DNA-directed RNA polymerase II subunit RPB1
• Protein Accession #: NP_000928.1
• Purification: Protein A purified
• Nucleotide Accession #: NM_000937.4
• Preservative: 0.05% Sodium Azide
• Gene Symbol: POLR2A
• Predicted Species Reactivity: Human
• Application: CHIP, WB, IF
• Image 1: Human Hela cells
  
  ChIP results obtained with the Aviva monoclonal antibody directed against Pol II S2p. ChIP assays were performed using human HeLa cells, the Aviva monoclonal antibody against Pol II S2p and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for 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 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
• Image 2: HelaS3 cells
  
  ChIP-seq results obtained with the Aviva monoclonal antibody directed against Pol II S2p r. ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 ug of the Aviva antibody against Pol II S2p 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 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome, and in a two genomic regions surrounding the GAPDH and ACTB positive control genes.
• Image 3: Pol IIS2p
  
  Cross reactivity of the Aviva monoclonal antibody directed against Pol IIS2p. To test the specificity an ELISA was performed using a serial dilution of the Aviva monoclonal antibody against Pol IIS2p. 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 S2 phosphorylation.
• Image 4: Hela cells
  
  Immunofluorescence using the Aviva monoclonal antibody directed against Pol II S2p. HeLa cells were stained with the Aviva antibody against Pol II S2p 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 S2p 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.
• Image 5: Human Hela cells
  
  Western blot analysis using the Aviva monoclonal antibody directed against Pol II S2p. Nuclear extracts (25 ug) from HeLa cells were analysed by Western blot using the Aviva monoclonal antibody against Pol II S2p diluted 1:1,000 in TBS-Tween containing 5% skim milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.