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MABE953

Sigma-Aldrich

Anti-phospho RNA Pol II (Ser2), clone 3E7C7 Antibody

clone 3E7C7, from rat

Synonym(s):

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

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

rat

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

3E7C7, monoclonal

species reactivity

monkey, canine, human, rat, mouse

technique(s)

ChIP: suitable (ChIP-seq)
ELISA: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgG2aκ

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

phosphorylation (pSer2)

Gene Information

human ... POLR2A(5430)

General description

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.

Immunogen

KLH-conjugated linear peptide corresponding to human RNA Pol II phosphorylated at Ser2.

Application

Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
This Anti-phospho RNA Pol II (Ser2) antibody, clone 3E7C7 is validated for use in western blotting, ChIP-seq, ELISA, ICC & ChIP for the detection of phospho RNA Pol II (Ser2).
Western Blotting Analysis: 0.5 µg/mL from a representative lot detected phospho RNA Pol II (Ser2) in 10 µg of HeLa nuclear extract.

Western Blotting: A representative lot detected phospho RNA Pol II (Ser2) 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 (Ser2) 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 (Ser2) 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 (Ser2) in a panel of unmodified and modified RNA Pol II (Ser2) 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 (Ser2) in HeLa cells (Prof. Taro Tachibana, Cell Engineering Corporation.).

Alexa Fluor is a registered trademark of Life Technologies.

Quality

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected phospho RNA Pol II (Ser2) in 10 µg of HeLa cell lysate.

Target description

~217 kDa observed

Physical form

Format: Purified
Protein G Purified
Purified rat monoclonal IgG2aκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

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

Other Notes

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

Legal Information

ALEXA FLUOR is a trademark of Life Technologies

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.

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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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 Research, 41, 54-62 (2013)
Jun Odawara et al.
BMC genomics, 12, 516-516 (2011-10-21)
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

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