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AbFlex® RNA Pol II antibody (rAb)

Aliases: RNA pol II, RNA Polymerase II
商品コード: 91151 Format: 100 µg ¥62,000 Buy Now
商品コード: 91152 Format: 10 µg ¥17,000 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A
Host:Mouse
Molecular Weight:217 kDa
Reactivity:Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated

for AbFlex RNA Pol II antibody (rAb)Application Notes


Validated Applications:
ChIP-Seq: 4 µg per ChIP
Western Blot: 0.1 – 1 µg/mL
Immunofluorescence (IF): 2 µg/mL.

for AbFlex RNA Pol II antibody (rAb)Immunogen

This RNA Pol II antibody was raised against a synthetic peptide YSPTSpPS corresponding to human RNA pol II. 

for AbFlex RNA Pol II antibody (rAb)Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 35 mM NaOAc, 0.035% sodium azide, 30% glycerol.

 
AbFlex<sup>®</sup> RNA Pol II antibody (rAb) tested by ChIP-Seq.

ChIP-Seq of AbFlex® RNA Pol II antibody (rAb).
ChIP was performed using AbFlex RNA Pol II recombinant antibody with 20 µg chromatin from the HT1080 cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 12 million sequence tags were mapped to identify RNA Pol II binding sites. The image shows binding across a region of chromosome 12.

AbFlex<sup>®</sup> RNA Pol II antibody (rAb) tested by Western blot.

Western Blot of AbFlex® RNA Pol II antibody (rAb).
Western Blot: 30 µg HeLa cell nuclear extract* was run on an SDS-PAGE gel and probed with AbFlex RNA Pol II (rAb) at 1 µg/mL.
*Nuclear extract prepared with high salt / sonication protocol. Contact Technical Service for additional info.
MW: 217 kDa

AbFlex<sup>®</sup> RNA Pol II antibody (rAb) tested by immunofluorescence.

Immunofluorescence stain of AbFlex® RNA Pol II antibody (rAb).
RNA Pol II was detected in human HeLa cells in a 2-step staining protocol using AbFlex RNA Pol II (rAb) at 2 µg/mL, followed by Anti-mouse IgG2a Dylight® 488.

for AbFlex RNA Pol II 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 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.

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 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 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 antibody (rAb).

for AbFlex RNA Pol II 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 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 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> RNA Pol II antibody (rAb) tested by ChIP-Seq.

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

AbFlex<sup>®</sup> RNA Pol II antibody (rAb) tested by Western blot.

Western blot of AbFlex® RNA Pol II antibody (rAb).
(Click image to enlarge and see details).

AbFlex<sup>®</sup> RNA Pol II antibody (rAb) tested by immunofluorescence.

Immunofluorescence stain of AbFlex® RNA Pol II antibody (rAb).
(Click image to enlarge and see details).