Anti-RNA polymerase II CTD repeat YSPTSPS [8WG16] antibody - ChIP Grade (ab817)
- Product nameAnti-RNA polymerase II CTD repeat YSPTSPS [8WG16] antibody - ChIP GradeSee all RNA polymerase II CTD repeat YSPTSPS primary antibodies ...
- DescriptionMouse monoclonal [8WG16] to RNA polymerase II CTD repeat YSPTSPS - ChIP Grade
- SpecificityThis antibody reacts with the highly conserved heptapeptide repeat of the largest subunit of eukaryotic RNA polymerase II.
- Tested applicationsCHIPseq, ICC, Purification, WB, ChIP, Flow Cyt, IP more details
- Species reactivityReacts with: Mouse, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Schizosaccharomyces pombe
Predicted to work with: a wide range of other species
Purified wheat germ RNA polymerase II.
- General notesThis 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.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
- Concentration information loading...
- PurityProtein G purified
- Clonality Monoclonal
- Clone number8WG16
- Epigenetics and Nuclear Signaling
- Polymerase associated factors
- Pol II Transcription
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.
|CHIPseq||CHIPseq: Use at an assay dependent dilution. PubMed: 19251593Use 2ug per 0.3ml of sonicated chromatin.|
|ICC||ICC: Use at an assay dependent dilution.|
|Purification||P: Use at an assay dependent dilution.|
|WB||WB: Use at an assay dependent dilution. Detects a band of approximately 217 kDa (predicted molecular weight: 217 kDa).|
|ChIP||ChIP: Use at an assay dependent dilution. See Abreview; the user recommends using anti-mouse IgG coated Dynabeads instead of Protein A to recover the precipitate).|
|Flow Cyt||Flow Cyt: Use 0.5µg for 106 cells.|
|IP||IP: Use at an assay dependent dilution.|
- FunctionDNA-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 similaritiesBelongs to the RNA polymerase beta' chain family.
modificationsThe 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 (By similarity). Following UV treatment, the elongating form of RNA polymerase II (RNA pol IIo) is ubiquitinated UV damage sites without leading to degradation: ubiquitination is facilitated by KIAA1530/UVSSA and promotes RNA pol IIo backtracking to allow access to the nucleotide excision repair machinery.
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 localizationNucleus.
- Entrez Gene: 177190 Caenorhabditis elegans
- Entrez Gene: 5430 Human
- Entrez Gene: 20020 Mouse
- Entrez Gene: 851415 Saccharomyces cerevisiae
- Omim: 180660 Human
- SwissProt: P18616 Arabidopsis thaliana
- SwissProt: P16356 Caenorhabditis elegans
- SwissProt: P24928 Human
- DNA directed RNA polymerase II A antibody
- DNA-directed RNA polymerase II largest subunit antibody
- DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd 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
- hRPB220 antibody
- hsRPB1 antibody
- POLR2 antibody
- Polr2a antibody
- POLRA antibody
- Polymerase (RNA) II (DNA directed) polypeptide A 220kDa antibody
- Polymerase (RNA) II (DNA directed) polypeptide A 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
- RPBh1 antibody
- RpIILS antibody
- RPO2 antibody
- RPOL2 antibody
Anti-RNA polymerase II CTD repeat YSPTSPS [8WG16] antibody - ChIP Grade images
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.
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.
Overlay histogram showing HeLa cells stained with ab817 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab817, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-RNA polymerase II CTD repeat YSPTSPS [8WG16] antibody - ChIP Grade (ab817)
This product has been referenced in:
- Caravaca JM et al. Bookmarking by specific and nonspecific binding of FoxA1 pioneer factor to mitotic chromosomes. Genes Dev 27:251-60 (2013). Read more (PubMed: 23355396) »
- Golla U et al. Sen1p contributes to genomic integrity by regulating expression of ribonucleotide reductase 1 (RNR1) in Saccharomyces cerevisiae. PLoS One 8:e64798 (2013). WB ; Saccharomyces cerevisiae . Read more (PubMed: 23741394) »