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Key Documents

17-620

Sigma-Aldrich

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

from mouse

Synonyme(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

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

Source biologique

mouse

Niveau de qualité

Clone

monoclonal

Espèces réactives

rat, Saccharomyces cerevisiae, mouse, human

Fabricant/nom de marque

ChIPAb+
Upstate®

Technique(s)

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

Isotype

IgG1

Numéro d'accès NCBI

Numéro d'accès UniProt

Conditions d'expédition

dry ice

Description générale

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

Spécificité

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

Immunogène

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).

Conditionnement

25 assays per set. ~1 μg per chromatin immunoprecipitation

Qualité

Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.

Description de la cible

210-220 kDa

Forme physique

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

Stockage et stabilité

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

Remarque sur l'analyse

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

Informations légales

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

Clause de non-responsabilité

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.

Code de la classe de stockage

10 - Combustible liquids


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Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Consulter la Bibliothèque de documents

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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)
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Mai N Tran et al.
The Journal of biological chemistry, 288(5), 3275-3288 (2012-12-15)
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Xiangbo Ruan et al.
The Journal of clinical investigation, 131(1) (2020-10-14)
A growing number of long noncoding RNAs (lncRNAs) have emerged as vital metabolic regulators. However, most human lncRNAs are nonconserved and highly tissue specific, vastly limiting our ability to identify human lncRNA metabolic regulators (hLMRs). In this study, we established

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