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Go-ChIP-Grade™ Purified anti-RNA Polymerase II Antibody

Pricing & Availability
Clone
CTD4H8 (See other available formats)
Regulatory Status
RUO
Other Names
DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II subunit B1, DNA-directed RNA polymerase II subunit A, RNA-directed RNA polymerase II subunit RPB1, DNA-directed RNA polymerase III largest subunit
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_CTD4H8_GoChIP_RNA_Polymerase_Antibody_071717
Chromatin Immunoprecipitations (ChIP) were performed with cross-linked chromatin samples from 4 X 106 of HeLa cells with either A)1:300 dilution of Go-ChIP-Grade™ Purified anti-RNA Polymerase II Antibody (clone CTD4H8, Cat. No. 904003) or B) equal amount of Purified Mouse IgG1, κ Isotype Control Antibody (Clone MG1-45, Cat. No. 401401) by using Go-ChIP-Grade™ Protein G Enzymatic Kit (Cat. No. 699904). The enriched DNA was purified and quantified by real-time qPCR using primers targeting human GAPDH gene region or α-Satellite repeats. The amount of immunoprecipitated DNA in each sample is represented as signal relative to total amount of input chromatin.
  • 1_CTD4H8_GoChIP_RNA_Polymerase_Antibody_071717
    Chromatin Immunoprecipitations (ChIP) were performed with cross-linked chromatin samples from 4 X 106 of HeLa cells with either A)1:300 dilution of Go-ChIP-Grade™ Purified anti-RNA Polymerase II Antibody (clone CTD4H8, Cat. No. 904003) or B) equal amount of Purified Mouse IgG1, κ Isotype Control Antibody (Clone MG1-45, Cat. No. 401401) by using Go-ChIP-Grade™ Protein G Enzymatic Kit (Cat. No. 699904). The enriched DNA was purified and quantified by real-time qPCR using primers targeting human GAPDH gene region or α-Satellite repeats. The amount of immunoprecipitated DNA in each sample is represented as signal relative to total amount of input chromatin.
  • 2_CTD4H8_GoChIP_RNA_PolymeraseII_Antibody_WB_071717
    Total lysates (15 µg protein) from HeLa (lane 1), Jurkat (lane 2) and Raw cells (lane 3) were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-RNA Polymerase II Antibody, clone CTD4H8 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-RNA Polymerase II Antibody or 1:5000 diluted Direct-Blot HRP anti-β-Actin Antibody, clone 2F1-1(lower). Lane M: Molecular weight ladder.
  • 3_CTD4H8_GoChIP_RNA_PolymeraseII_Antibody_WB_071717
    Western blot probed with anti-RNA Polymerase II antibody (clone CTD4H8) Lane 1 : Molecular weight ladder; Lane 2: BSA negative control; Lane 3 : HeLa cell lysate.
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904004 100 µg $259
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Description

RPB1 is the catalytic and largest component of RNA polymerase II, which synthesizes mRNA precursors and many functional non-coding RNAs. It forms the polymerase active center together with RPB2, the second largest subunit. Polymerase II (Pol II) is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relatively 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 DNA template strand of the promoter is positioned within the central active site cleft of Pol II. Then, a bridging helix emanates from RPB1 and crosses the cleft near the catalytic site, which acts as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations during each neuocleotide addition. This promotes translocation of Pol II. Pol II moves on the template during transcription elongation. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II's largest subunit (RPB1), which serves as a platform for assembling factors that regulate transcription initiation, elongation, termination, and mRNA processing. It can act as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, being able to conform as both a replicate and transcriptase for the viral RNA circular genome.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Reported Reactivity
Other species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
The immunogen used was a peptide containing 10 repeats of the synthetic peptide YSPTSPS using chemically synthesized phospho-ser5.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

ChIP - Quality tested
WB - Verified
ICC - Reported in the literature, not verified in house 
Recommended Usage

Each lot of this antibody is quality control tested by ChIP Assay. The suggested dilution for ChIP application is 1:300-1:500 by volume. For Western blotting the suggested working dilution is 1:500 by volume. For immunofluorescent staining, the suggested dilution is 1:500 by volume.

It is recommended that the reagent to be titrated for optimal performance before each experiment.

Application Notes

This antibody is effective in immunoblotting, immunoprecipitation and ELISA. It can also be used in chromatin immunoprecipitation assays.

The antibody CTD4H8 recognizes the C-terminal repeat of the largest subunit of RNA polymerase II from HeLa and S. cerevisiae cells. This antibody recognizes both the phosphorylated and unphosphorylated forms of the RNA polymerase II.

Based on sequence identify, this clone is liable to react with a broad range of species.

Clone CTD4H8 is also known as 4H8 or CTD 4H8.

Predicted MW is ~ 217 kD, Observed MW is on WB gel is ~240 kD

Application References

(PubMed link indicates BioLegend citation)
  1. Kristjuhan A, et al. 2002. Mol. Cell. 10:925.
  2. Charos AE, et al. 2012. Genome Res. 22:1668. (ChIP)
  3. Rafalska-Metcalf IU, et al. 2010. PLoS One 5:e10272. (ICC) PubMed
RRID
AB_2715775 (BioLegend Cat. No. 904004)

Antigen Details

Biology Area
Cell Biology, Signal Transduction, Transcription Factors
Molecular Family
Nuclear Markers
Gene ID
5430 View all products for this Gene ID
UniProt
View information about RNA Polymerase II on UniProt.org
Go To Top Version: 6    Revision Date: 09/21/2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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