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Analysis Note

ControlIncluded negative control antibody mouse IgG and control primers specific for human GAPDH.

Application

Western Blot Analysis:
3T3 nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-RNA polymerase II (0.1 µg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Research CategoryEpigenetics & Nuclear Function

RNA Pol II ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers. The antibody ChIP is used for chromatin immunoprecipitation of RNA Pol II .

Research Sub CategoryChromatin Biology

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ RNA Pol II set includes the RNA Pol II antibody, the negative control antibody (mouse IgG), and qPCR primers flanking the human GAPDH promoter, yielding a 166 bp product. The RNA Pol II and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of RNA Pol II associated chromatin.

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

Immunogen

The RNA Pol II antibody is made against a peptide corresponding to the C-terminal domain of RNA Pol II.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

25 assays per set. ~1 µg per chromatin immunoprecipitation

Physical form

Anti-RNA Pol II (mouse monoclonal IgG1,purified). One vial containing 25 µg of purified antibody in 25 µL volume. Store at -20°C.

Normal Mouse IgG. One vial containing 25 ug of mouse IgG in 25 µL volume. Store at -20°C.

Control Primers p21. One vial containing 75 µL of 5 µM of each primer specific for a region of the human GAPDH promoter. Store at -20°C.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG
CGA

Protein G Purified

Format: Purified

Quality

Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.

Specificity

Recognizes RNA Pol II

Other species predicted to cross-react based upon sequence conservation are rat and yeast.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Target description

210-220 kDa