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Merck
CN

17-620

Sigma-Aldrich

ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set

from mouse

Synonym(s):

Chip Antibody and primer set, DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II, RNA polymerase II ChIP

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

Key Documents

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biological source

mouse

Quality Level

100

clone

monoclonal

species reactivity

rat, Saccharomyces cerevisiae, mouse, human

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
immunofluorescence: suitable
immunoprecipitation (IP): suitable

isotype

IgG1

NCBI accession no.

NP_000928

UniProt accession no.

P24928

shipped in

dry ice

General description

210-220 kDa
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.

Application

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 Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
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).

Biochem/physiol Actions

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

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
Format: Purified
Protein G Purified

Preparation Note

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

Analysis Note

Control
Included negative control antibody mouse IgG and control primers specific for human GAPDH.
Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.

Legal Information

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

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.

Storage Class Code

10 - Combustible liquids

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Pistoni, M; Shiue, L; Cline, MS; Bortolanza, S; Neguembor, MV; Xynos, A; Ares, M; Gabellini, D
PLoS Genetics null
Johanna Meier-Soelch et al.
Frontiers in immunology, 9, 775-775 (2018-05-15)
The potent proinflammatory cytokine interleukin (IL)-1 triggers gene expression through the NF-κB signaling pathway. Here, we investigated the cofactor requirements of strongly regulated IL-1 target genes whose expression is impaired in p65 NF-κB-deficient murine embryonic fibroblasts. By two independent small-hairpin
Xiaomei Liu et al.
Journal of neuroinflammation, 18(1), 108-108 (2021-05-12)
Interleukin 9 (IL-9), produced mainly by T helper 9 (Th9) cells, has been recognized as an important regulator in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Astrocytes respond to IL-9 and reactive astrocytes always associate with
Mai N Tran et al.
The Journal of biological chemistry, 288(5), 3275-3288 (2012-12-15)
Epithelial-mesenchymal transition (EMT) is a physiological process that plays important roles in tumor metastasis, "stemness," and drug resistance. EMT is typically characterized by the loss of the epithelial marker E-cadherin and increased expression of EMT-associated transcriptional repressors, including ZEB1 and
Xiangbo Ruan et al.
Nature communications, 11(1), 45-45 (2020-01-04)
Unlike protein-coding genes, the majority of human long non-coding RNAs (lncRNAs) are considered non-conserved. Although lncRNAs have been shown to function in diverse pathophysiological processes in mice, it remains largely unknown whether human lncRNAs have such in vivo functions. Here
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