RNA Pol II CTD phospho Tyr1 antibody (mAb)

Clone: 3D12
Aliases: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
Catalog No: 61383 Format: 100 µg $455 Buy
Catalog No: 61384 Format: 10 µg $110 Buy
Catalog No: 61983 Format: 50 µg $265 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1
Purification:
Protein G Chromatography
Host:
Rat
Molecular Weight:
210 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated Western Blot Validated Immunoprecipitation Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes


Validated Applications:
WB*: 1 – 2 µg/ml

*Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot. In addition, we recommend the addition of 0.05% Tween 20 to all blocking solutions to reduce background. Individual optimization may be required.

Published Applications:
ChIP
IP
WB
IF/ICC

See references for more information.

Also available as an AbFlex® engineered recombinant antibody.  For details on the corresponding AbFlex Recombinant Antibody, see Catalog No. 91219.

Immunogen

This antibody was raised against a synthetic peptide containing the RNA Pol II heptad repeat consensus sequence phosphorylated at tyrosine 1.

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide.

References

 

RNA pol II CTD phospho Tyr1 mAb tested by Western blot.
20 µg of HeLa cell nuclear extract was run on SDS-PAGE and probed with antibody at 2 µg/ml.

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 sites within the heptapeptide repeat, at serine 2, serine 5, serine 7 and tyrosine 1. RNA pol II Tyrosine 1 phosphorylation is found during elongation and appears to activate transcription by suppressing RNA pol II termination.

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 Tyr1 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