RNA pol II CTD phospho Ser7 antibody (mAb)

RRID: AB_2687452
Clone: 4E12
Aliases: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
Catalog No: 61087 Format: 100 µg $455 Buy
Catalog No: 61088 Format: 10 µg $110 Buy
Catalog No: 61987 Format: 50 µg $265 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1
Purification:
Protein G Chromatography
Host:
Rat
Molecular Weight:
240 kDa
Reactivity:
Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunoprecipitation Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes


Validated Applications:
ChIP: 5-10 µg per ChIP
ChIP-Seq: 5-10 µg each

Published Applications:
ChIP
IP
ICC/IF
WB
See references for more information. Individual optimization may be required.

Immunogen

This RNA pol II CTD phospho Ser7 antibody was raised against a peptide containing the RNA pol II CTD sequence phosphorylated at serine 7.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 

RNA pol II CTD phospho Ser7 antibody (mAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 2.3 million HL-60 cells and 4 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 20 million sequence tags were mapped to identify Pol II phospho Ser7 binding. The image shows a 350,000 base pair region on chromosome 1 with the expected enrichment at gene promoters.

Background

RNA pol II (RNA polymerase II) is responsible for synthesizing messenger RNA in eukaryotes. RNA pol II contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, RNA pol II, in combination with several other polymerase subunits, form the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA.

During the transcription cycle, the CTD of the large subunit of RNA pol II is reversibly phosphorylated. RNA pol II containing unphosphorylated CTD is recruited to the promoter, whereas the hyperphosphorylated CTD form is involved in active transcription. Phosphorylation occurs at two sites within the heptapeptide repeat, at serine 2, serine 5 and serine 7. RNA pol II Serine 7 phosphorylation is confined to promoter regions and is necessary for the initiation of transcription.

Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for RNA Pol II CTD phospho Ser7 antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with RNA pol II CTD phospho Ser7 antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • WB = Western Blotting