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Products:Epigenetics and Nuclear Signaling >> Transcription >> Polymerase associated factors >> Pol II Transcription
Overview
Product name
Anti-RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade
See all RNA polymerase II CTD repeat YSPTSPS products (17) ...
Description
Rabbit polyclonal to RNA polymerase II CTD repeat YSPTSPS - ChIP Grade
Tested applications
WB, IHC-P, ICC/IF, ChIPmore details
Cross reactivity
Reacts with
Mouse, Human
Immunogen
Synthetic peptide of Human RNA polymerase II CTD repeat YSPTSPS.
(Peptide available as ab17564.)
Positive control
This antibody gave a positive control in both HeLa and HEK293 whole cell lysates.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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
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ChIP - RNA polymerase II CTD repeat YSPTSPS antibody (ab26721)
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Immunocytochemistry/ Immunofluorescence - RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
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Applications
Show applications key
Our Abpromise guarantee covers the use of ab26721 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
- 1 Image
IHC-P
Application notes
IHC-P: Use at a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP: Use 5-10µg for 25µg of chromatin
ICC/IF: Use at a concentration of 1 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 220 kDa (predicted molecular weight: 220 kDa). Can be blocked with RNA polymerase II CTD repeat YSPTSPS peptide (ab17564).
Not yet tested in other applications.
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. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphataes, 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).
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
see all
Database links
- Entrez Gene: 5430 Human
- Entrez Gene: 20020 Mouse
- SwissProt: P24928 Human
- SwissProt: P08775 Mouse
- Unigene: 270017 Human
- Unigene: 16533 Mouse
Anti-RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade images:
ChIP - RNA polymerase II CTD repeat YSPTSPS antibody (ab26721)
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 5µg of ab26721 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the inactive AFM and F8 promoters, the GAPDH promoter (active) and over the g-Actin gene (active). Schematic diagram of the g-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
Immunocytochemistry/ Immunofluorescence - RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
ab26721 (1/200) staining RNA polymerase II in assyncchonous HeLa cells (green). cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.
Image courtesy of an abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
IHC image of RNA polymerase staining in human lymphoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab26721, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunocytochemistry/ Immunofluorescence - RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
ICC/IF image of ab26721 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab26721, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HEK 293, HepG2 and MCF7 cells.
Western blot - RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with RNA polymerase II CTD repeat YSPTSPS peptide (ab17564) at 1 µg/ml
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with RNA polymerase II CTD repeat YSPTSPS peptide (ab17564) at 1 µg/ml
Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5 (ab18488) at 1 µg/ml
Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5 (ab18488) at 1 µg/ml
Lane 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2 (ab12793) at 1 µg/ml
Lane 8 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2 (ab12793) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 220 kDa
Observed band size : 220 kDa
Exposure time : 1 minute
References for Anti-RNA polymerase II CTD repeat YSPTSPS antibody - ChIP Grade (ab26721)
This product has been referenced in:
- Thorne JLet al. Epigenetic control of a VDR-governed feed-forward loop that regulates p21(waf1/cip1) expression and function in non-malignant prostate cells. Nucleic Acids Res 39:2045-56 (2011). ChIP; Human.Read more (PubMed: 21088000) »
- Wellberg Eet al. The bHLH/PAS transcription factor singleminded 2s promotes mammary gland lactogenic differentiation. Development 137:945-52 (2010). ChIP; Mouse.Read more (PubMed: 20150276) »
See all 2 publications for this product
Publishing research using ab26721? 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"