Anti-phospho RNA Pol II (Ser5), clone 1H4B6 Antibody

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.

KLH-conjugated linear peptide corresponding to human RNA PoI II phosphorylated at Ser5.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Transcription Factors

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).

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).

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.

~217 kDa observed

Protein G Purified

Format: Purified

Purified rat monoclonal IgG2bκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.