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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- ELISA [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- NBP2-59219 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Mouse Monoclonal RNA Polymerase II/POLR2A Antibody
- Antibody type
- Monoclonal
- Description
- Protein A purified.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Vial size
- 50 ug
- Storage
- Store at -20C. Avoid freeze-thaw cycles.
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Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Western Blot: RNA Polymerase II/POLR2A [p Ser5] Antibody (C.15200007) [NBP2-59219] - Whole cell extracts (40 ug) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the 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 (kDa) is shown on the left.
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Western Blot: RNA Polymerase II/POLR2A [p Ser5] Antibody (C.15200007) [NBP2-59219] - Nuclear extracts (25 ug) from HeLa cells were analysed by Western blot using the 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 (kDa) is shown on the left.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- ELISA: RNA Polymerase II/POLR2A [p Ser5] Antibody (C.15200007) [NBP2-59219] - Cross reactivity of the antibody directed against Pol IIS5p. To test the specificity an ELISA was performed using a serial dilution of the antibody against Pol IIS5p. The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure shows the specificity of the antibody for the S5 phosphorylation.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: RNA Polymerase II/POLR2A [p Ser5] Antibody (C.15200007) [NBP2-59219] - ChIP assays were performed using human HeLa cells, the antibody against Pol II S5p and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, 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 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 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
