AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb)

RRID: AB_2793780
Aliases: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
Catalog No: 91115 Format: 100 µg $385 Buy
Catalog No: 91116 Format: 10 µg $80 Buy

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Antibody Type:
Recombinant
Isotype:
IgG2a
Purification:
Ni-NTA
Host:
Mouse
Molecular Weight:
240 kDa
Reactivity:
Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Enzyme-linked Immunosorbent Assay Validated Immunocytochemistry Validated

Application Notes


Validated Applications:
ChIP-Seq: 4 µg each
ICC/IF: 1 - 2 µg / mL
WB: 0.5 - 2 µg/mL
Bead-based ELISA: 0.1 - 5 µg/ml

Published Applications:
IF*
Proximity Ligation Assay (PLA)*

*Oqani, RK, Lin, T, et. al. (2019), "Iws1 and Spt6 Regulate Trimethylation of Histone H3 on Lysine 36 through Akt Signaling and are Essential for Mouse Embryonic Genome Activation." Sci. Rep., Mar 7;9(1):3831.

AbFlex® recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental clone, see Catalog No. 61083.

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 Ser2 antibody was raised against a peptide containing the RNA pol II CTD sequence phosphorylated at serine 2.

Buffer

Purified IgG in 200 mM Hepes pH 7.5, 100 mm NaCl, 50 mM NaOAc, 30% glycerol and 0.035% sodium azide.

References

 

AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg chromatin from an accute large B-cell lymphoma cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 19 million sequence tags were mapped to identify RNA pol II phospho Ser2 binding. The image shows binding across a region of chromosome 17. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb) tested by immunofluorescence.
HeLa cell stained with 2µg /mL of AbFlexTM RNA pol II CTD phospho Ser2 antibody (rAb) followed by anti-mouse-IgG-488.

AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with RNA pol II CTD phospho Ser2 antibody (rAb) (0.5 µg/ml dilution).

AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb) tested by Luminex bead-based specificity analysis.
Luminex bead-based specificity analysis was used to confirm the specificity of AbFlexTM RNA pol II CTD phospho Ser2 antibody (rAb) antibody for RNA pol II CTD peptides. RNA pol II peptides were conjugated to MagPlex Luminex beads and incubated with various amounts of AbFlexTM RNA pol II CTD phospho Ser2 antibody (rAb). Peptide-bound antibody was detected with anti-mouse IgG-Phycoerythrin and read in a Luminex instrument.

Luminex® is a registered trademark of Luminex Corporation.

Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-Tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.

AbFlex® RNA pol II CTD phospho Ser2 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.

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 phosphorylated at serine 2 is enriched over the gene body and is associated with 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 Ser2 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 AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb).

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