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MABE954
Sigma-AldrichAnti-phospho RNA Pol II (Ser5), clone 1H4B6 Antibody
This Anti-phospho RNA Pol II (Ser5) antibody, clone 1H4B6 is validated for use in western blotting, ChIP-seq, ICC, ELISA & ChIP for the detection of phospho RNA Pol II (Ser5).
More>>This Anti-phospho RNA Pol II (Ser5) antibody, clone 1H4B6 is validated for use in western blotting, ChIP-seq, ICC, ELISA & ChIP for the detection of phospho RNA Pol II (Ser5). Less<<
MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Anti-phospho RNA Pol II (Ser5), clone 1H4B6 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
Background Information
RNA polymerase II (RNAPII or Pol II) is a multi-subunit enzyme responsible for the transcription of transcription of DNA into RNA from protein-coding genes. Transcription initiation requires recruitment of the complete transcription machinery to a promoter via solicitation by activators and chromatin remodeling factors. RNAPII can coordinate 10 to 14 subunits. This complex interacts with the promoter regions of genes and a variety of elements and transcription factors. The DNA binding domain of the polymerase is a groove where TFIIB orients the DNA for unwinding and transcription. 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 (Ser5) antibody, clone 1H4B6 is validated for use in western blotting, ChIP-seq, ICC, ELISA & ChIP for the detection of phospho RNA Pol II (Ser5).
Key Applications
Western Blotting
ChIP-seq
Immunocytochemistry
Enzyme Immunoassay (ELISA)
Chromatin Immunoprecipitation (ChIP)
Application Notes
Western Blotting Analysis: 0.5 µg/mL from a representative lot detected phospho RNA Pol II (Ser5) in 10 µg of HeLa nuclear extract.
Western Blotting: A representative lot detected phospho RNA Pol II (Ser5) in 10 µg of HeLa cell lysate (Odawara, J., et al. (2011). BMC Genomics. 12:516.).
Chromatin Immunoprecipitation Analysis: A representative lot from an independent laboratory immunoprecipitated phospho RNA Pol II (Ser5) in HeLa cell lysate (Odawara, J., et al. (2011). BMC Genomics. 12:516.; Maehara, K., et al. (2013). Nucleic Acids Res. 41(1):54-62.).
Chromatin Immunoprecipitation-Sequencing Analysis: A representative lot from an independent laboratory immunoprecipitated phospho RNA Pol II (Ser5) in HeLa cell lysate (Odawara, J., et al. (2011). BMC Genomics. 12:516.; Maehara, K., et al. (2013). Nucleic Acids Res. 41(1):54-62.).
ELISA Analysis: A representative lot from an independent laboratory detected phospho RNA Pol II (Ser5) in a panel of unmodified and modified RNA Pol II (Ser5) proteins (Prof. Taro Tachibana, Cell Engineering Corporation).
Immunocytochemistry Analysis: A 1:500 dilution from a representative lot from an independent laboratory detected phospho RNA Pol II (Ser5) in HeLa cells (Prof. Taro Tachibana, Cell Engineering Corporation).
Biological Information
Immunogen
KLH-conjugated linear peptide corresponding to human RNA PoI II phosphorylated at Ser5.
Clone
1H4B6
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Evaluated by Western Blotting in HeLa cell lysate.
Western Blotting Analysis: 0.5 µg/mL of this antibody detected phospho RNA Pol II (Ser5) 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
100 µg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number
GTIN
MABE954
04053252970627
Documentation
Anti-phospho RNA Pol II (Ser5), clone 1H4B6 Antibody SDS
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.
The classification of mRNA expression levels by the phosphorylation state of RNAPII CTD based on a combined genome-wide approach. Odawara, Jun, et al. BMC Genomics, 12: 516 (2011)
2011
Cellular function is regulated by the balance of stringently regulated amounts of mRNA. Previous reports revealed that RNA polymerase II (RNAPII), which transcribes mRNA, can be classified into the pausing state and the active transcription state according to the phosphorylation state of RPB1, the catalytic subunit of RNAPII. However, genome-wide association between mRNA expression level and the phosphorylation state of RNAPII is unclear. While the functional importance of pausing genes is clear, such as in mouse Embryonic Stem cells for differentiation, understanding this association is critical for distinguishing pausing genes from active transcribing genes in expression profiling data, such as microarrays and RNAseq. Therefore, we examined the correlation between the phosphorylation of RNAPII and mRNA expression levels using a combined analysis by ChIPseq and RNAseq.
