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AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb)

Aliases: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
商品コード: 91119 Format: 100 µg ¥62,000 Buy Now
商品コード: 91120 Format: 10 µg ¥17,000 Buy Now

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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

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)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

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)Immunogen

This RNA pol II CTD phospho Ser5 antibody was raised against a peptide containing the RNA pol II CTD sequence phosphorylated at serine 5.

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)Buffer

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

AbFlex™ recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones.  For details on the parental clone, see Catalog No. 61085.

 
AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by ChIP-Seq.

AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 20 µg of chromatin from HL60 cells and 4 µg of AbFlex® RNA pol II CTD phospho Ser5 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 20 million sequence tags were mapped to identify RNA pol II phospho Ser5 binding. The image shows binding across a region of chromosome 4.

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by immunofluorescence.

AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb) tested by immunofluorescence.
HeLa cell stained with 2µg /mL of AbFlexTM RNA pol II CTD phospho Ser5 antibody (rAb) followed by anti-mouse-IgG-488.

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by Western blot.

AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with AbFlexTM RNA pol II CTD phospho Ser5 antibody (0.5 µg/ml dilution).

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by ELISA.

AbFlex® RNA pol II CTD phospho Ser5 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 Ser5 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 Ser5 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.

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)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 Ser5 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 IIcontains 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 5 phosphorylation is confined to promoter regions and is necessary for the initiation of transcription.

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)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.

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 Ser5 antibody (rAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb).

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for AbFlex RNA pol II CTD phospho Ser5 antibody (rAb)Guarantee

This product is guaranteed for 6 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;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by ChIP-Seq.

ChIP-Seq of AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by immunofluorescence.

Immunofluorescence stain of AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by Western blot.

Western blot of AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> RNA pol II CTD phospho Ser5 antibody (rAb) tested by ELISA.

ELISA of AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb).
(Click image to enlarge and see details.)