製品: EZH2 Antibody
カタログ: AF5150
タンパク質の説明: Rabbit polyclonal antibody to EZH2
アプリケーション: WB IHC
反応性: Human, Mouse, Rat
予測: Pig, Zebrafish, Bovine, Horse, Rabbit, Dog, Chicken, Xenopus
分子量: 85kD, 100kD; 85kD(Calculated).
ユニプロット: Q15910
RRID: AB_2837636

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製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Zebrafish(90%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(92%)
クローナリティ:
Polyclonal
特異性:
EZH2 Antibody detects endogenous levels of total EZH2.
RRID:
AB_2837636
引用形式: Affinity Biosciences Cat# AF5150, RRID:AB_2837636.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ/展開

Enhancer of zeste 2; enhancer of zeste 2 polycomb repressive complex 2 subunit; Enhancer of zeste homolog 2 (Drosophila); Enhancer of zeste homolog 2; Enhancer of zeste, Drosophila, homolog 2; ENX 1; Enx 1h; ENX-1; ENX1; Enx1h; EZH 2; EZH1; EZH2; EZH2_HUMAN; EZH2b; Histone-lysine N-methyltransferase EZH2; KMT 6; KMT6; KMT6A; Lysine N-methyltransferase 6; MGC9169; WVS; WVS2;

免疫原

免疫原:
Uniprot:
遺伝子(ID):
発現特異性:
Q15910 EZH2_HUMAN:

Expressed in many tissues. Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis.

タンパク質の説明:
Polycomb 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.
タンパク質配列:
MGQTGKKSEKGPVCWRKRVKSEYMRLRQLKRFRRADEVKSMFSSNRQKILERTEILNQEWKQRRIQPVHILTSVSSLRGTRECSVTSDLDFPTQVIPLKTLNAVASVPIMYSWSPLQQNFMVEDETVLHNIPYMGDEVLDQDGTFIEELIKNYDGKVHGDRECGFINDEIFVELVNALGQYNDDDDDDDGDDPEEREEKQKDLEDHRDDKESRPPRKFPSDKIFEAISSMFPDKGTAEELKEKYKELTEQQLPGALPPECTPNIDGPNAKSVQREQSLHSFHTLFCRRCFKYDCFLHPFHATPNTYKRKNTETALDNKPCGPQCYQHLEGAKEFAAALTAERIKTPPKRPGGRRRGRLPNNSSRPSTPTINVLESKDTDSDREAGTETGGENNDKEEEEKKDETSSSSEANSRCQTPIKMKPNIEPPENVEWSGAEASMFRVLIGTYYDNFCAIARLIGTKTCRQVYEFRVKESSIIAPAPAEDVDTPPRKKKRKHRLWAAHCRKIQLKKDGSSNHVYNYQPCDHPRQPCDSSCPCVIAQNFCEKFCQCSSECQNRFPGCRCKAQCNTKQCPCYLAVRECDPDLCLTCGAADHWDSKNVSCKNCSIQRGSKKHLLLAPSDVAGWGIFIKDPVQKNEFISEYCGEIISQDEADRRGKVYDKYMCSFLFNLNNDFVVDATRKGNKIRFANHSVNPNCYAKVMMVNGDHRIGIFAKRAIQTGEELFFDYRYSQADALKYVGIEREMEIP

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Dog
100
Chicken
100
Rabbit
100
Xenopus
92
Zebrafish
90
Sheep
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q15910 基板として

Site PTM Type Enzyme
K7 Ubiquitination
S8 Phosphorylation
K10 Ubiquitination
S21 Phosphorylation P31749 (AKT1)
Y23 Phosphorylation
K39 Ubiquitination
K61 Ubiquitination
S75 O-Glycosylation
S76 O-Glycosylation
S76 Phosphorylation
S212 Phosphorylation
S220 Phosphorylation
K234 Acetylation
K234 Ubiquitination
Y244 Phosphorylation
T261 Phosphorylation
S277 Phosphorylation
T302 Phosphorylation
T305 Phosphorylation
T311 Phosphorylation
K318 Ubiquitination
T339 Phosphorylation
T345 Phosphorylation P24941 (CDK2) , P06493 (CDK1)
K348 Acetylation
S362 Phosphorylation
S363 Phosphorylation P49841 (GSK3B)
S366 Phosphorylation
T367 Phosphorylation P49841 (GSK3B) , Q16539 (MAPK14)
T369 Phosphorylation
S375 Phosphorylation
S380 Phosphorylation
T388 Phosphorylation
S405 Phosphorylation
S406 Phosphorylation
S408 Phosphorylation
S412 Phosphorylation
T416 Phosphorylation P24941 (CDK2)
T460 Phosphorylation
K461 Ubiquitination
K472 Ubiquitination
S474 Phosphorylation
S475 Phosphorylation
T487 Phosphorylation P06493 (CDK1)
K505 Methylation
K509 Methylation
K510 Methylation
K569 Ubiquitination
K597 Ubiquitination
K602 Ubiquitination
K629 Ubiquitination
K634 Ubiquitination
Y641 Phosphorylation O60674 (JAK2)
S690 Phosphorylation
Y696 Phosphorylation
K713 Ubiquitination
T718 Phosphorylation
S729 Phosphorylation
K735 Methylation
K735 Ubiquitination

研究背景

機能:

Polycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) 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. Displays a preference for substrates with less methylation, loses activity when progressively more methyl groups are incorporated into H3K27, H3K27me0 > H3K27me1 > H3K27me2. 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. 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. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4 and the nuclear receptor RORA. Regulates the circadian clock via histone methylation at the promoter of the circadian genes. Essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK-ARNTL/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.

PTMs:

Phosphorylated by AKT1. Phosphorylation by AKT1 reduces methyltransferase activity. Phosphorylation at Thr-345 by CDK1 and CDK2 promotes maintenance of H3K27me3 levels at EZH2-target loci, thus leading to epigenetic gene silencing.

Sumoylated.

Glycosylated: O-GlcNAcylation at Ser-75 by OGT increases stability of EZH2 and facilitates the formation of H3K27me3 by the PRC2/EED-EZH2 complex.

細胞の位置付け:

Nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Expressed in many tissues. Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis.

サブユニット構造:

Binds ATRX via the SET domain (Probable). Component of the PRC2/EED-EZH2 complex, which includes EED, EZH2, SUZ12, RBBP4 and RBBP7 and possibly AEBP2. The minimum components required for methyltransferase activity of the PRC2/EED-EZH2 complex are EED, EZH2 and SUZ12. The PRC2 complex may also interact with DNMT1, DNMT3A, DNMT3B and PHF1 via the EZH2 subunit and with SIRT1 via the SUZ12 subunit. Interacts with HDAC1 and HDAC2. Interacts with PRAME. Interacts with CDYL. Interacts with CLOCK, ARNTL/BMAL1 and CRY1 (By similarity). Interacts with DNMT3L; the interaction is direct (By similarity). Interacts with EZHIP; the interaction blocks EZH2 methyltransferase activity.

タンパク質ファミリー:

Belongs to the class V-like SAM-binding methyltransferase superfamily. Histone-lysine methyltransferase family. EZ subfamily.

研究領域

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Metabolism > Amino acid metabolism > Lysine degradation.

参考文献

1). EZH2 deficiency attenuates Treg differentiation in rheumatoid arthritis. JOURNAL OF AUTOIMMUNITY, 2020 (PubMed: 31952907) [IF=12.8]

Application: WB    Species: Human    Sample: CD4+ T cells

Fig. 1. EZH2 expression in PBMC and CD4+ T cells from RA and HC. (A) EZH2 mRNA expression in PBMC, CD4+ T cells, CD14+ monocytes and CD19+ B cells from HCs and RA patients (n = 22). (B) EZH2 protein expression in peripheral CD4+cells from HCs and RA patients (n = 5). (C) EZH2 protein expression in peripheral CD4+ CD45RA+ naïve T cells from HCs and RA patients (n = 20). (**: p < 0.01, ****: p < 0.0001).

2). N6-methyladenosine-associated prognostic pseudogenes contribute to predicting immunotherapy benefits and therapeutic agents in head and neck squamous cell carcinoma. Theranostics, 2023 (PubMed: 36438489) [IF=12.4]

Application: WB    Species: Human    Sample: ARO and Tca8113 cell

Figure 4 m6A-associated pseudogene can regulate targeted immune-involved genes via miRNAs. (A) Sankey showing pseudogenes together with binding miRNAs and target genes with | R | ≥ 0.3 and P < 0.05 were used to construct the pseudogene-miRNA-target gene regulatory networks by subtypes of oncogene pseudogene PDIA3P1 and tumor-suppressor pseudogene RRN3P3. The column on the left represented pseudogenes, which are located at the cores of the networks. The column in the middle and the column on the right stand for binding miRNAs and target genes, respectively. (B-G) Experimental validation of PDIA3P1 affects the expression of AKT1 via miR-34a-5p in ARO and Tca8113 cell lines. (B) Relative gene expression of PDIA3P1 after PDIA3P1 knockdown using siRNA. (C) Relative gene expression of AKT1 after PDIA3P1 knockdown using siRNA. (D) Western blot comparing the protein levels of AKT1 in control and PDIA3P1 knockdown cells. (E) The relative expression of AKT1 at different time points after transcription inhibition in control and PDIA3P1 knockdown cells respectively. Error bars represent standard errors. (F) The relative expression of PDIA3P1 and AKT1 after adding control inhibitor versus miR-34a-5p inhibitor. (G) Western blot comparing the protein levels of AKT1 after adding control inhibitor versus miR-34a-5p inhibitor. (H-M) Experimental validation of RRN3P3 affects the expression of EZH2 via miR-26b-5p in ARO and Tca8113 cell lines. (H) Relative gene expression of RRN3P3 after RRN3P3 knockdown using siRNA. (I) Relative gene expression of EZH2 after RRN3P3 knockdown using siRNA. (J) The relative expression of EZH2 at different time points after transcription inhibition in control and RRN3P3 knockdown cells respectively. Error bars represent standard errors. (K) Western blot comparing the protein levels of EZH2 in control and RRN3P3 knockdown cells. (L) The relative expression of RRN3P3 and EZH2 after adding control inhibitor versus miR-26b-5p inhibitor. (M) Western blot comparing the protein levels of EZH2 after adding control inhibitor versus miR-26b-5p inhibitor. (N) UCSC genome browser tracks m6A-seq and m6A-LAIC-seq data indicating m6A peaks and m6A levels of oncogene pseudogene PDIA3P1 and tumor-suppressor pseudogene RRN3P3. Read-coverage tracks of input, m6A-negative, and m6A-positive fractions of m6A-LAIC-seq shown along with overlay tracks of m6A-seq (cyan for input and red for RIP; predicted m6A sites in m6A peaks are indicated by arrows). Read coverage (y-axis) of m6A negative and m6A positive are normalized as previously described 17 to reflect the calculated m6A levels (i.e., equal signals in m6A positive (eluate) versus m6A negative (supernatant) suggest m6A levels of 50%), while input and IP tracks of m6A-seq are shown for optimal viewing at the top panel. * P< 0.05; ** P< 0.01; *** P< 0.001 (two-tailed t-test). (O) The m6A methylation level of the pseudogenes at specific modification sites (Chr1:146650342, GG(m6A)CA on PDIA3P1; Chr16:22431201, GG(m6A)CG on RRN3P3) using SELECT in control and METTL3 knockdown ARO and Tca8113 cell.

3). E3 ubiquitin ligase STUB1 affects the mTORC1 pathway through p62 and participates in regulating the differentiation of follicular helper T cells in rheumatoid arthritis. Clinical immunology (Orlando, Fla.), 2023 (PubMed: 37604355) [IF=8.6]

4). TNIK drives castration-resistant prostate cancer via phosphorylating EGFR. iScience, 2024 (PubMed: 38226156) [IF=5.8]

5). miR‑101a‑3p overexpression prevents acetylcholine‑CaCl2‑induced atrial fibrillation in rats via reduction of atrial tissue fibrosis, involving inhibition of EZH2. Molecular Medicine Reports, 2021 (PubMed: 34435649) [IF=3.4]

Application: IHC    Species: rat    Sample: atrial

Figure 3.| Pathological staining for evaluating the severity of AF in rats. (C) Immunohistochemistry assay was utilized to examine EZH2 expression in atrial tissues. Scale bar, 50 µm.

Application: WB    Species: Rat    Sample: atrial tissues

Figure 1. Expression levels of miR-101a-3p and EZH2 in atrial tissues of rats. The rats were anesthetized with an intraperitoneal injection of pentobarbital sodium (50 mg/kg), fixed, intubated and injected with LV-NC or LV-miR-101a-3p. After injection for 48 h, all rats were anesthetized. Then, 1 ml/kg acetylcholine-CaCl2 mixture via the tail vein was injected into rats daily for 7 days. At day 8, rats were anesthetized, injected with Ach-CaCl2 and sacrificed. The atrial tissues of heart were collected for following experiments. (A) Experimental protocol in rats. (B) miR-101a-3p expression was detected via reverse transcription-quantitative PCR. (C) Protein expression level of EZH2 was measured via western blotting. β-actin was used as an internal reference. The results are presented as the mean ± standard deviation (n=6). ##P<0.01 and ###P<0.001 vs. sham group; ***P<0.001 vs. AF + LV-NV group. LV-NC, lentivirus negative control; AF, atrial fibrillation; miR, microRNA; EZH2, enhancer of zeste 2 homolog 2; SD, Sprague-Dawley.

6). Evaluation of Immunohistochemical Expression of Enhancer of Zeste Homolog 2 (EZH2) and Its Association With Clinicopathological Variables in Carcinoma Cervix. Cureus, 2022 (PubMed: 37131568) [IF=1.2]

Application: IHC    Species: Human    Sample:

Figure 3 A: Showing invasive nests of basaloid cells in a case of basaloid SCC (H&E x4). B: Corresponding IHC in the same case showing strong nuclear positivity of EZH2 (Score 9) (IHC EZH2 x20). C: Showing papillae with fibrovascular core and lined by squamous epithelium in papillary SCC (H&E x10). D Corresponding IHC showing strong nuclear positivity of EZH2 (Score 9) (IHC EZH2, x20).

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