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Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

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Overview

Product name

Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade
See all RNA polymerase II CTD repeat YSPTSPS products (17) ...

Description

Mouse monoclonal [8WG16] to RNA polymerase II CTD repeat YSPTSPS - ChIP Grade

Specificity

This antibody reacts with the highly conserved heptapeptide repeat of the largest subunit of eukaryotic RNA polymerase II.

Tested applications

CHIPseq, ICC, Purification, WB, ChIP, IPmore details

Cross reactivity

Reacts with

Mouse, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Schizosaccharomyces pombe

Predicted to work with

a wide range of other species

Immunogen

Purified wheat germ RNA polymerase II.

General notes

This antibody may be used to detect unproteolyzed RNA polymerase II or to inhibit specific transcription from class II promoters. This is a polyol-responsive antibody and can be used in gentle purifications of RNA polymerase II from from wheat germ, calf thymus, and yeast and is likely to purify RNA polymerase II from most eukaryotic organisms. It inhibits promoter-directed transcription, but it does not inhibit elongation in the nonspecific transcription assay.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.02% Sodium Azide
Constituents: PBS

Concentration

Concentration information loading...

Purity

Protein G purified

Clonality

Monoclonal

Clone number

8WG16

Isotype

IgG2a

Research areas

Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Transcription >> RNA polymerase
Epigenetics and Nuclear Signaling >> Transcription >> Polymerase associated factors >> Pol II Transcription >> Polymerase
Epigenetics and Nuclear Signaling >> Transcription >> Polymerase associated factors >> Pol II Transcription

  • Western blot - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)Western blot - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817) image (enlarge)

  • ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817) image (enlarge)

  • ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab817 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Target

Function

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 a 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.

Sequence similarities

Belongs to the RNA polymerase beta' chain family.

Post-translational
modifications

The tandem 7 residues repeats in the C-terminal domain (CTD) can be highly phosphorylated. The phosphorylation activates Pol II. Phosphorylation occurs mainly at residues 'Ser-2' and 'Ser-5' of the heptapeptide repeat and is mediated, at least, by CDK7 and CDK9. CDK7 phosphorylation of POLR2A associated with DNA promotes transcription initiation by triggering dissociation from DNA. Phosphorylation also takes place at 'Ser-7' of the heptapepdtide repeat, which is required for efficient transcription of snRNA genes and processing of the transcripts. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphatases, and a "CTD code" that specifies the position of Pol II within the transcription cycle has been proposed.
Dephosphorylated by the protein phosphatase CTDSP1.
Ubiquitinated by WWP2 leading to proteasomal degradation.
Methylated at Arg-1810 by CARM1. Methylation occurs only when the CTD is hypophosphorylated, and phosphorylation at Ser-1805 and Ser-1808 prevent methylation (in vitro). It is assumed that methylation occurs prior to phosphorylation and transcription initiation. CTD methylation may facilitate the expression of select RNAs.

Cellular localization

Nucleus.

Target information above from: UniProt accessionP24928 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

Alternative names

  • DNA directed RNA polymerase II A antibody
  • DNA-directed RNA polymerase II largest subunit antibody
  • DNA-directed RNA polymerase II subunit A antibody
  • DNA-directed RNA polymerase II subunit RPB1 antibody
  • DNA-directed RNA polymerase III largest subunit antibody
  • Polr2a antibody
  • RNA pol II CTD antibody
  • RNA polymerase II subunit B1 antibody
  • RNA-directed RNA polymerase II subunit RPB1 antibody
  • RPB1 antibody
  • RPB1_HUMAN antibody
see all

Database links

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Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade images:

  Western blot - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

Western blot - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817) at 1/10 dilution

Lane 1 : 1ug non-phosphorylated recombinant RNA polymerase II protein
Lane 2 : 1ug phosphorylated recombinant RNA polymerase II protein


Predicted band size : 217 kDa
Observed band size : 217 kDa

  ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

Various regions across the Actin2/7 loci were tested for the presence of RNA polymerase II CTD repeat YSPTSPS. A nuclear lysate from Arabidopsis thaliana seedlings was crosslinked using 1% formaldehyde for 30 seconds.  The ChIP was performed with 0.1 µg of ab817 per µg of chromatin; incubated together for 16 hours at 4°C.  The immunoprecipitated DNA was quantified by Real-Time PCR.  The bottom panel indicates the positive (ab817) and negative controls (no antibody) at region B3.

This image is courtesy of an Abreview submitted by Dr Szymon Swiezewski

See Abreview

  ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

ChIP - RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

Chromatin was prepared from nuclear lysate of the human MCF7 breast epithelial adenocarcinoma cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. The primary antibody was used in concentration of 0.2 µg/µg chromatin and incubated with the sample for 16 hours at 4°C in SDS, DOC, TritonX-100, EDTA, HEPES, NaCl. The immunoprecipitated DNA was quantified by real time PCR. Ct values were converted to DNA copy numbers using a standard curve in the Q-PCR step. The number of binding events detected for each test reaction was then calculated by taking into account the DNA copy number, cell equivalents of chromatin used in the ChIP and PCR, and primer pair amplification efficiency.

This image is a courtesy of Genpathway Inc

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References for Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817)

This product has been referenced in:

  • Li MD  et al. O-GlcNAc Transferase Is Involved in Glucocorticoid Receptor-mediated Transrepression. J Biol Chem 287:12904-12 (2012).Read more (PubMed: 22371499) »
  • Bergmann JH  et al. Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore. EMBO J 30:328-40 (2011). ChIP; Human.Read more (PubMed: 21157429) »

See all 49 publications for this product

Publishing research using ab817? Please let us know so that we can cite the reference in this datasheet

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"