RNA pol II CTD phospho Ser5 antibody (pAb)

RRID: AB_2793198
Aliases: RNA Polymerase II Carboxy Terminal Doman phosphoserine 5, RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
Catalog No: 39233 Format: 100 µl $455 Buy
Catalog No: 39234 Format: 10 µl $110 Buy

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Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
210 kDa
Reactivity:
Human, Mouse, Wide Range Predicted

Applications

ChIP Validated Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes


Validated Applications:
ChIP: 10 µl per ChIP
ICC/IF: 1:250 - 1:500 dilution
WB: 1:500 - 1:2,000 dilution

Published Applications:
ChIP
IF

For RNA pol II CTD phospho Ser5, we also offer AbFlex® RNA pol II CTD phospho Ser5 Recombinant Antibody (rAb).  For details, see Catalog No. 91119.

Immunogen

This RNA pol II CTD phospho Ser5 antibody was raised against synthetic peptide containing the RNA Pol II heptad repeat consensus sequence phosphorylated at serine 5.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39749) of this antibody that was purified by Protein A Chromatography is also available.

References

 

RNA pol II CTD phospho Ser5 antibody tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 µl of RNA pol II CTD phospho Ser5 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the PABPC1 gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

RNA pol II CTD phospho Ser5 antibody tested by immunofluorescence.
Top image: HeLa cells stained with RNA pol II CTD phospho Ser5 antibody at a 1:1,000 dilution. Middle: Same cells stained with DAPI. Bottom: Merge of both images.

RNA pol II CTD phospho Ser5 antibody tested by Western blot.
Nuclear extract of HeLa cells (20 µg) probed with RNA pol II CTD phospho Ser5 antibody at a 1:2,000 dilution.

RNA pol II CTD phospho Ser5 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of RNA pol II CTD phospho Ser5 antibody for phospho-Ser5 of the RNA Pol II C-terminal domain heptad repeat. Modified and unmodified peptides were spotted onto PVDF and probed with the antibody at a dilution of 1:1,000. Decreasing amounts of peptide were spotted in each row.
    Lane 1: Peptide phosphorylated at CTD repeat serine 2.
    Lane 2: Unmodified CTD repeat serine 2 peptide.
    Lane 3: Peptide phosphorylated at CTD repeat serine 5.
    Lane 4: Unmodified CTD repeat serine 5 peptide.

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 5 and Serine 2. RNA pol II Serine 5 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 Ser5 antibody.

qPCR Primers

Active Motif's Human Positive Control Primer Set ACTB-2 (Cat. No. 71005) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Positive Control Primer Set Actb-2 (Cat. No. 71017) and Mouse Negative Control Primer Set 1 (Cat. No. 71011) have been validated for both qPCR and endpoint PCR.

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 Ser5 antibody (pAb).

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