RNA Pol II CTD phospho Thr4 antibody (pAb)

RRID: AB_2793588
Aliases: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
Catalog No: 61307 Format: 100 µg $445 Buy
Catalog No: 61308 Format: 10 µg $110 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Molecular Weight:
240 kDa
Reactivity:
Human

Applications

Western Blot Validated

Application Notes


Validated Applications:
WB: 1 - 2 µg/ml dilution

The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection by Western blot. Individual optimization may be required.

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.

Immunogen

This RNA Pol II CTD phospho Thr4 antibody was raised against a peptide containing the RNA Pol II CTD sequence phosphorylated at threonine 4.

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 

RNA pol II CTD phospho Thr4 antibody pAb tested by Western blot.
Nuclear extract of HeLa cells (40 µg) probed with RNA pol II CTD phospho Thr4 antibody at a dilution of 2 µg/ml.

RNA Pol II CTD phospho Thr4 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of RNA Pol II CTD phospho Thr4 pAb. Peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at a dilution of 1 µg/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
    Lane 1: Phospho Thr4 of RNA Pol II CTD peptide.
    Lane 2: Unmodified Thr4 of RNA Pol II CTD peptide.
    Lane 3: Phospho Ser2 of RNA Pol II CTD peptide.
    Lane 4: Phospho Ser5 of RNA Pol II CTD peptide.

Background

RNA pol II (RNA polymerase II) is a complex of 12 proteins that participates in transcription, mRNA processing, and transcription-coupled DNA repair. RNA polymerase II is distinguished by its large carboxyl-terminal repeat domain (CTD), composed of repeats of the consensus heptapeptide Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7. Polo-like kinase 3 (plk3) has been reported to phosphorylate Thr4 which may be required for transcriptional elongation.

Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for RNA Pol II CTD phospho Thr4 antibody.

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