General description
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.
~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.
Immunogen
KLH-conjugated linear peptide corresponding to human phospho RNA Pol II (Ser7).
Application
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.).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
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).
Physical form
Format: Purified
Protein G Purified
Purified rat monoclonal IgG2bκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Preparation Note
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
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.
Disclaimer
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.
| biological source | rat |
| Quality Segment | 100 |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 3D4A12, monoclonal |
| species reactivity | canine, human, monkey, rat, mouse |
| concentration | 1 mg/mL |
| technique(s) | ChIP: suitable (ChIP-seq),ELISA: suitable,immunocytochemistry: suitable,western blot: suitable |
| isotype | IgG2bκ |
| NCBI accession no. | NP_000928 |
| UniProt accession no. | P24928 |
| shipped in | wet ice |
| target post-translational modification | phosphorylation (pSer7) |
| Gene Information | human ... POLR2A(5430) |
