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Antibodies

AbFlex® EZH2 antibody (rAb)

Catalog No: 91159 Format: 100 µg $375 Buy Now
Catalog No: 91160 Format: 10 µg $75 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A Chromatography
Host:Mouse
Reactivity:Human

Applications

Immunoprecipitation Validated

for AbFlex EZH2 antibody (rAb)Application Notes


Validated Applications:
IP: 4 µg/ml per IP

*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. In addition, we recommend adding 500mM NaCl to the IP wash buffer for best performance.

for AbFlex EZH2 antibody (rAb)Immunogen

This antibody was raised against a recombinant protein corresponding to amino acids 353-451 of human EZH2.

for AbFlex EZH2 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> EZH2 antibody (rAb) tested by immunoprecipitation.

AbFlex® EZH2 antibody (rAb) tested by Immunoprecipitation.
HeLa cell nuclear extract* was immunoprecipitated with 4 µg of AbFlex EZH2 antibody (lane 1) or control IgG2a (lane 2). Immunoprecipitates were run on SDS-PAGE and analyzed by Western blot using EZH2 polyclonal antibody (Cat. No. 39999) at 2 µg/ml, followed by incubation with Anti-Rabbit light chain secondary antibody.

for AbFlex EZH2 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® EZH2 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.

EZH2 – (Enhancer of Zeste homolog 2, also designated Enx1 and SETDB1) is a human homolog of the Drosophila Enhancer of Zeste protein. It contains a SET domain that catalyzes the mono-, di- and trimethylation of Histone H3 Lys27. EZH2, EED and Suz12 are present in the PRC protein complexes that function as mediators of epigenetic transcriptional silencing. Polycomb group proteins repress gene expression by binding to chromatin and locally altering chromatin structure. Deregulation of EZH2 is linked to a variety of cancers.

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® EZH2 antibody (rAb).

for AbFlex EZH2 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 EZH2 antibody (rAb)Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

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® EZH2 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> EZH2 antibody (rAb) tested by immunoprecipitation.

Immunoprecipitation of AbFlex® EZH2 antibody (rAb).
(Click image to enlarge and see details.)