EZH2 Antibody

Affinity Purified Rabbit Polyclonal Antibody (Pab)

All lanes: Anti-EZH2 Antibody at 1:2000 dilution Lane 1: Mouse spleen lysate Lane 2: Mouse testis lysate Lysates/proteins at 20 µg per lane. Secondary: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ASP1615) at 1/15000 dilution. Observed band size: 85 KDa Blocking/Dilution buffer: 5% NFDM/TBST.
Overlay histogram showing Hela cells stained with AP2512D (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP2512D, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NA168821) at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Overlay histogram showing Hela cells stained with AP2512D (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP2512D, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NA168821) at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized Hela (Human cervical epithelial adenocarcinoma cell line) cells labeling EZH2 with AP2512d at 1/25 dilution, followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG (1583138) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear staining on Hela cell line. Cytoplasmic actin is detected with Alexa Fluor® 555 conjugated with Phalloidin (OB16636430) at 1/100 dilution (red).
All lanes : Anti-EZH2 Antibody at 1:2000 dilution Lane 1: Jurkat whole cell lysates Lane 2: MDA-MB-231 whole cell lysates Lane 3: MDA-MB-468 whole cell lysates Lane 4: T47D whole cell lysates Lane 5: mouse spleen lysates Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution Predicted band size : 85 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot analysis of lysates from MDA-MB-468, SW620, T47D cell line, mouse spleen, mouse testis tissue (from left to right), using EZH2 Antibody(Cat. #AP2512d). AP2512d was diluted at 1:1000 at each lane. A goat anti-rabbit IgG H&L(HRP) at 1:10000 dilution was used as the secondary antibody. Lysates at 20ug per lane.
Formalin-fixed and paraffin-embedded human testis tissue reacted with EZH2 Antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IF, FC, E
Primary Accession	Q15910
Reactivity	Human, Mouse
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	85363 Da
Antigen Region	1-296 aa

Additional Information
Gene ID	2146
Other Names	Histone-lysine N-methyltransferase EZH2, ENX-1, Enhancer of zeste homolog 2, Lysine N-methyltransferase 6, EZH2, KMT6
Target/Specificity	This EZH2 antibody is generated from rabbits immunized with a recombinant fragment (N-term) protein from human EZH2.
Dilution	WB~~1:1000 IHC-P~~1:100~500 IF~~1:25 FC~~1:25 E~~Use at an assay dependent concentration.
Format	Purified polyclonal antibody supplied in PBS with 0.05% (V/V) Proclin 300. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	EZH2 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	EZH2 (HGNC:3527)
Synonyms	KMT6
Function	Catalytic subunit of the PRC2/EED-EZH2 complex, a Polycomb group (PcG) complex that methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene (PubMed:14532106, PubMed:15225548, PubMed:15385962, PubMed:16618801, PubMed:16936726, PubMed:17344414, PubMed:22323599, PubMed:24474760, PubMed:26581166, PubMed:30026490, PubMed:30923826). Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively (PubMed:15231737, PubMed:17210787, PubMed:18285464, PubMed:22323599, PubMed:30923826). Displays a preference for substrates with less methylation, loses activity when progressively more methyl groups are incorporated into H3K27, H3K27me0 > H3K27me1 > H3K27me2 (PubMed:22323599, PubMed:30923826). Compared to EZH1-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 (PubMed:19026781). The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems (PubMed:16357870, PubMed:17200670). Genes repressed by the PRC2/EED- EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes (PubMed:16179254, PubMed:18086877, PubMed:20935635). EZH2 can also methylate non-histone proteins such as the transcription factor GATA4 and the nuclear receptor RORA (PubMed:23063525). Regulates the circadian clock via histone methylation at the promoter of the circadian genes (PubMed:16717091). Essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK- BMAL1 heterodimer; involved in the di and trimethylation of 'Lys-27' of histone H3 on PER1/2 promoters which is necessary for the CRY1/2 proteins to inhibit transcription (By similarity).
Cellular Location	Nucleus. Note=Localizes to the inactive X chromosome in trophoblast stem cells. {ECO:0000250\|UniProtKB:Q61188}
Tissue Location	In the ovary, expressed in primordial follicles and oocytes and also in external follicle cells (at protein level) (PubMed:31451685). Expressed in many tissues (PubMed:14532106) Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis (PubMed:14532106)