• USA Home
  • 17-620 - ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set

EMAIL THIS PAGE TO A FRIEND
17-620 Sigma-Aldrich

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

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 .

Synonym: DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II

  •  eCl@ss 32160702

  •  NACRES NA.32

Purchase

Properties

Related Categories Antibodies, Antibodies for Epigenetics and Nuclear Signaling, ChIP Validated Antibodies, Primary Antibodies
biological source   mouse
clone   monoclonal
species reactivity   Saccharomyces cerevisiae, mouse, mouse, rat, human, human
mfr. no.   ChIPAb+
  Upstate®
application(s)   ChIP: suitable
  immunofluorescence: suitable
  immunoprecipitation (IP): suitable
isotype   IgG1
NCBI accession no.   NP_000928
UniProt accession no.   P24928
shipped in   dry ice

Description

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

Specificity

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

Recognizes RNA Pol II

Immunogen

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

Application

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).

Packaging

25 assays per set. ~1 μg per chromatin immunoprecipitation

Quality

Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.

Target description

210-220 kDa

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

Storage and Stability

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.

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.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Related Content

Antibody Explorer | Buy Primary Antibodies & Secondary Antibodies

Primary, Secondary and Recombinant Monoclonal Antibodies Providing highly cited primary and secondary antibodies, we have you covered for your ELISA, western blot, immunohistochemistry or other assay...
Keywords: Alzheimer Disease, Cancer, Chromatin immunoprecipitation, Diagnostic, Enzyme-linked immunosorbent assay, Flow cytometry, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Neuroscience, Western blot

Peer-Reviewed Papers
15

References

Related Products

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?