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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 (phospho S5) antibody
See all RNA polymerase II CTD repeat YSPTSPS products (17) ...
Description
Rabbit polyclonal to RNA polymerase II CTD repeat YSPTSPS (phospho S5)
Specificity
ab52208 detects endogenous levels of RNA polymerase II only when phosphorylated at serine 1619.
Tested applications
WB, ICC/IF, ELISA, IHC-Pmore details
Cross reactivity
Reacts with
Human
Predicted to work with
Mouse, Rat
Immunogen
Synthesized phosphopeptide derived from human RNA polymerase II CTD repeat YSPTSPS around the phosphorylation site of serine 5 (P-T-SP-P-S).
Positive control
Human breast carcinoma tissue; extracts from COS7 cells.
Properties
Form
Liquid
Storage instructions
Store at -20°C. Stable for 12 months at -20°C
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Clonality
Polyclonal
Isotype
IgG
Research areas
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
Applications
Show applications key
Our Abpromise guarantee covers the use of ab52208 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: 1/500 - 1/1000.Detects a band of approximately 217 kDa (predicted molecular weight: 217 kDa).
ICC/IF: Use a concentration of 1 µg/ml
ELISA
ELISA: 1/10000ELISA: 1/10000
IHC-P: 1/50 - 1/100.
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
- MGC75453 antibody
- B220 antibody
- DNA directed RNA polymerase II A antibody
see all
Database links
- Entrez Gene: 5430 Human
- Entrez Gene: 20020 Mouse
- Entrez Gene: 363633 Rat
- SwissProt: P24928 Human
- SwissProt: P08775 Mouse
- Unigene: 270017 Human
- Unigene: 16533 Mouse
Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody images:
Immunohistochemistry (Paraffin-embedded sections) - RNA polymerase II (phospho S1619) antibody (ab52208)
This image shows human breast carcinoma tissue stained with ab52208 at a dilution of 1/50 - 1/100. Right hand image: tissue treated with immunising peptide; left hand image: untreated.
Western blot - RNA polymerase II (phospho S1619) antibody (ab52208)
All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab52208) at 1/500 dilution
Lane 1 : Extracts from COS7 cells treated
with EGF (200ng/ml, 30min) with no immunising peptide
Lane 2 : Extracts from COS7 cells treated
with EGF (200ng/ml, 30min) with immunising peptide
Predicted band size : 217 kDa
Observed band size : 217 kDa
Immunocytochemistry/ Immunofluorescence-RNA polymerase II (phospho S1619) antibody(ab52208)
ICC/IF image of ab52208 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52208, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab52208)
ab52208 has not yet been referenced specifically in any publications.
Publishing research using ab52208? 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"