EZH2 antibody (pAb)

RRID: AB_2614956
Aliases: WVS, ENX1, EZH1, KMT6, WVS2, ENX-1, EZH2b, KMT6A
Catalog No: 39901 Format: 100 µl $445 Buy
Catalog No: 39002 Format: 50 µl $260 Buy
Catalog No: 39902 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
96 kDa
Reactivity:
Human, Mouse

Applications

ChIP Validated ChIP-Seq Validated Immunoprecipitation Validated Immunohistochemistry Validated

Application Notes


Validated Applications:
ChIP: 5 µl per ChIP
ChIP-Seq: 5 µl each
IHC (FFPE): 1:1000 dilution

This antibody has not been validated in WB, but a WB-validated antibody is available for this target: 39933.

Published Applications:
ChIP
ChIP-Seq
Proxity Ligation Assay (PLA)
IP
See references for more information. Individual optimization may be required.

Immunogen

This EZH2 antibody was raised against a recombinant fusion protein corresponding to amino acids 1-370 of mouse EZH2.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 61641) of this antibody that was purified by Protein A Chromatography is also available.

References

Serebryannyy, L.A., et. al. 2019. "HiPLA: High-throughput imaging proximity ligation assay." Methods. Mar 15;157:80-87. (Proximity Ligation Assay)

(Proximity Ligation Assay)

 

EZH2 antibody (pAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 4 million SUDHL6 B cell lymphoma cells and 5 µl of EZH2 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 25 million sequence tags were mapped to identify EZH2 binding sites. The image on the left shows EZH2 binding at the 5’ end of the CCND2 gene. The image on the right shows EZH2 binding across the repressed HOX gene cluster.

EZH2 antibody (pAb) tested by ChIP.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 4 million SUDHL6 B cell lymphoma cells and 5 µl of EZH2 antibody. ChIP DNA was used in qPCR with the negative control primer pair or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

EZH2 antibody (pAb) tested by Immunohistochemistry
Nuclear staining pattern is detected in Formalin-fixed, paraffin-embedded tissue sections from human substantia nigra (midbrain). Top Panel: 5-mC antibody at 1:1000 dilution. Bottom Panel: No primary antibody (2nd step antibody alone)

Background

EZH2(Enhancer of Zeste homolog 2) is a human homolog of the Drosophila Polycomb-group protein Enhancer of Zeste protein. It contains a SET domain that catalyzes the methylation of histone H3 at lysine 27 (Histone H3 Lys27 monomethylated, dimethylated and trimethylated). Polycomb-group proteins repress gene expression by binding to chromatin and locally altering chromatin structure. EZH2, BMI-1 and Suz12 are present in the PRC2 and PRC3 protein complexes that function as mediators of epigenetic transcriptional silencing. EZH2 associates with the embryonic ectoderm development protein, the VAV1 oncoprotein, and the X-linked nuclear protein. EZH2 may play a role in the hematopoietic and central nervous systems. Deregulation of EZH2 is linked to a variety of cancers.

Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for EZH2 antibody.

qPCR Primers

Active Motif's Human Positive Control Primer Set MYT1 (Cat. No. 71007) and Human Negative Control Primer Set 3 (Cat. No. 71023) 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 Zc3h13 (Cat. No. 71022) 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 EZH2 antibody (pAb).

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.

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