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MABE955
Sigma-AldrichAnti-phospho RNA Pol II (Ser7), clone 3D4A12 Antibody
This Anti-phospho RNA Pol II (Ser7) antibody, clone 3D4A12 is validated for use in western blotting, ChIP-seq, ChIP, ICC & ELISA for the detection of phospho RNA Pol II (Ser7).
More>>This Anti-phospho RNA Pol II (Ser7) antibody, clone 3D4A12 is validated for use in western blotting, ChIP-seq, ChIP, ICC & ELISA for the detection of phospho RNA Pol II (Ser7). Less<<
MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Anti-phospho RNA Pol II (Ser7), clone 3D4A12 Antibody
Alternate Names
DNA-directed RNA polymerase II subunit RPB1
RNA polymerase II subunit B1
DNA-directed RNA polymerase II subunit A
DNA-directed RNA polymerase III largest subunit
RNA-directed RNA polymerase II subunit RPB1
RNAPII
RNA PolII
RNAPIIS7p
Background Information
As the study of the RNA polymerase II (RNAPII) transcription complex expands, many researchers are interested in using immunoblots to detect RNAPII in various protein preparations and column fractions. HeLa cell RNAPII contains 10 subunits, the largest possessing a heptapeptide repeat (PTSPSYS) in the C-terminal domain (CTD). This large subunit cycles, during the course of transcription, through nonphosphorylated (RNAPIIA) and hyperphosphorylated (RNAPIIO) forms. RNAPIIA associates with the basal transcription complex and becomes phosphorylated to the RNAPIIO form before the synthesis of the first phosphodiester bond. The timing of these events suggests that this phosphorylation event may function as a regulatory point, and there is considerable interest in identifying components of the transcription complex with CTD kinase activity.
References
Product Information
Format
Purified
Presentation
Purified rat monoclonal IgG2bκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
This Anti-phospho RNA Pol II (Ser7) antibody, clone 3D4A12 is validated for use in western blotting, ChIP-seq, ChIP, ICC & ELISA for the detection of phospho RNA Pol II (Ser7).
Key Applications
Western Blotting
ChIP-seq
Chromatin Immunoprecipitation (ChIP)
Immunocytochemistry
ELISA
Application Notes
Chromatin Immunoprecipitation-Sequencing Analysis: A representative lot from an independent laboratory immunoprecipitated phospho RNA Pol II (Ser7) from HeLa cell lysate (Maehara, K., et al. (2013). Nucleic Acids Res. 41(1):54-62.).
Chromatin Immunoprecipitation Analysis: A representative lot from an independent laboratory immunoprecipitated phospho RNA Pol II (Ser7) from HeLa cell lysate (Maehara, K., et al. (2013). Nucleic Acids Res. 41(1):54-62.).
Western Blotting Analysis: 1 µg/mL from a representative lot detected phospho RNA Pol II (Ser7) in HeLa (lane 1), COS1, L929, NRK, MDCK, and DM4 whole cell extracts (Prof. Taro Tachibana, Cell Engineering Corporation.).
Immunocytochemistry Analysis: A 1:500 dilution from representative lot detected phospho RNA Pol II (Ser7) in HeLa cells (Prof. Taro Tachibana, Cell Engineering Corporation.).
ELISA Analysis: A representative lot detected phospho RNA Pol II (Ser7) in a panel of unmodified and modified RNA Pol II peptides (Prof. Taro Tachibana, Cell Engineering Corporation.).
Biological Information
Immunogen
KLH-conjugated linear peptide corresponding to human phospho RNA Pol II (Ser7).
~270 kDa observed. Uncharacterized band(s) may be observed in some cell lysates. This protein may be observed at higher molecular weights due to post-translational modification.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in HeLa cell lysate.
Western Blotting Analysis: 4 µg/mL of this antibody detected phospho RNA Pol II (Ser7) in 10 µg of HeLa cell lysate.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at 2-8°C from date of receipt.
Packaging Information
Material Size
200 µg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number
GTIN
MABE955
04053252970634
Documentation
Anti-phospho RNA Pol II (Ser7), clone 3D4A12 Antibody Certificates of Analysis
A co-localization model of paired ChIP-seq data using a large ENCODE data set enables comparison of multiple samples. Maehara, Kazumitsu, et al. Nucleic Acids Res., 41: 54-62 (2013)
2013
Deep sequencing approaches, such as chromatin immunoprecipitation by sequencing (ChIP-seq), have been successful in detecting transcription factor-binding sites and histone modification in the whole genome. An approach for comparing two different ChIP-seq data would be beneficial for predicting unknown functions of a factor. We propose a model to represent co-localization of two different ChIP-seq data. We showed that a meaningful overlapping signal and a meaningless background signal can be separated by this model. We applied this model to compare ChIP-seq data of RNA polymerase II C-terminal domain (CTD) serine 2 phosphorylation with a large amount of peak-called data, including ChIP-seq and other deep sequencing data in the Encyclopedia of DNA Elements (ENCODE) project, and then extracted factors that were related to RNA polymerase II CTD serine 2 in HeLa cells. We further analyzed RNA polymerase II CTD serine 7 phosphorylation, of which their function is still unclear in HeLa cells. Our results were characterized by the similarity of localization for transcription factor/histone modification in the ENCODE data set, and this suggests that our model is appropriate for understanding ChIP-seq data for factors where their function is unknown.
Although ERBB2 amplification and overexpression is correlated with poor outcome in breast cancer, the molecular mechanisms underlying the aggressive nature of these tumors has not been fully elucidated. To investigate this further, we have used a transgenic mouse model of ErbB2-driven tumor progression (ErbB2(KI) model) that recapitulates clinically relevant events, including selective amplification of the core erbB2 amplicon. By comparing the transcriptional profiles of ErbB2(KI) mammary tumors and human ERBB2-positive breast cancers, we show that ErbB2(KI) tumors possess molecular features of the basal subtype of ERBB2-positive human breast cancer, including activation of canonical β-catenin signaling. Inhibition of β-catenin-dependent signaling in ErbB2(KI)-derived tumor cells using RNA interference impaired tumor initiation and metastasis. Furthermore, treatment of ErbB2(KI) or human ERBB2-overexpressing tumor cells with a selective β-catenin/CBP inhibitor significantly decreased proliferation and ErbB2 expression. Collectively, our data indicate that ERBB2-mediated breast cancer progression requires β-catenin signaling and can be therapeutically targeted by selective β-catenin/CBP inhibitors.
