WB: Hek293 whole cell lysate; EZH2 recombinant
ICC: mouse embryonic stem cells
IP: U20S cells transfected with 4ug of Myc-EZH2
The Drosophila protein Enhancer of Zeste is a member of the Polycomb group, which maintains homeotic gene repression and is thought to control gene expression by regulating chromatin. EZH2 is thought to perform a similar role.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Primary antibody notesThe Drosophila protein Enhancer of Zeste is a member of the Polycomb group, which maintains homeotic gene repression and is thought to control gene expression by regulating chromatin. EZH2 is thought to perform a similar role.
Use at an assay dependent concentration. PubMed: 19571010
FunctionPolycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' and 'Lys-27' of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes.
Tissue specificityExpressed in many tissues. Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis.
Sequence similaritiesBelongs to the histone-lysine methyltransferase family. EZ subfamily. Contains 1 SET domain.
Developmental stageExpression decreases during senescence of embryonic fibroblasts (HEFs). Expression peaks at the G1/S phase boundary.
Post-translational modificationsPhosphorylated by AKT1. Phosphorylation by AKT1 reduces methyltransferase activity.
Immunocytochemistry/ Immunofluorescence - Anti-KMT6 / EZH2 antibody - ChIP Grade (ab3748)Image courtesy of an anonymous Abreview.
ab3748 staining KMT6/EZH2 in the HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehye permeabilized with 0.2% Triton X-100 in PBS and blocked with 2% BSA for 45 minutes at RT. Samples were incubated with primary antibody (1/2550 in PBS + 2% BSA) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1:10000).
ChIP - Anti-KMT6 / EZH2 antibody - ChIP Grade (ab3748)This image is courtesy of an anonymous Abreview
ChIP analysis using ab3748 binding KMT6/EZH2 in rat Hippocampal primary Neurons. Cells were cross-linked for 10 minutes with 1% formaldehyde. Samples were incubated with primary antibody (1:100) for 16 hours at 4°C. Protein binding was detected using real-time PCR.
Positive control: HoxA1, a PolyComb repressed gene in Hippocampus.
Negative Control:b-act, an active gene, and normal Rabbit IgG.
U20S cells were transfected with 4ug of Myc-EZH2. WB reprobe using 0.4ug/ml in TBS milk 2%, BSA 0.5%.
NB. ab3748 did not detect endogenous EZH2.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - KMT6 / EZH2 antibody (ab3748)Image courtesy of an anonymous Abreview.
ab3748 staining KMT6 / EZH2 in murine hippocampus tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using 10mM sodium citrate. Samples were then permeabilized using 0.3% Triton, blocked with 3% BSA for 30 minutes at 22°C and then incubated with ab3748 at a 1/3000 dilution for 16 hours at 4°C. The secondary used was a biotin conjugated horse polyclonal, used at a 1/2000 dilution. ABC method.
Immunocytochemistry/ Immunofluorescence - Anti-KMT6 / EZH2 antibody - ChIP Grade (ab3748)This image is a courtesy of Anonymous Abreview
ab3748 staining KMT6 / EZH2 in human U-2 OS cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton ×100 and blocking with 1% BSA was done for 1 hour at RT. Samples were incubated with primary antibody (1/100) for 2 hours. An undiluted Alexa Fluor®594-conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.
ChIP - Anti-KMT6 / EZH2 antibody (ab3748)This image is courtesy of an anonymous Abreview
ChIP analysis of Human glioma cells using ab3748 to bind KMT6 / EZH2. Chromatin was obtained by cross-linking with 1% formaldehyde for 10 minutes and incubated with primary antibody (1/250) for 16 hours at 4°C. Protein binding was detected using semi-quantitive PCR.