Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Ezh2 (D2C9) XP® Rabbit mAb #5246

histone methyltransferase   HMT   sc-25383  

No. Size Price
5246S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
5246T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
5246 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 98 Rabbit IgG
IP 1:300
IHC-P 1:50
F 1:200
IF-F 1:100
IF-IC 1:200
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-F=Immunofluorescence (Frozen), IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

Ezh2 (D2C9) XP® Rabbit mAb detects endogenous levels of total Ezh2 protein.

Ezh2 (D2C9) XP® Rabbit mAb兔单抗能够检测内源性Ezh2总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg354 of human Ezh2 protein.

通过合成的与人源Ezh2蛋白Arg354位点相应的多肽片段去免疫动物从而制备出此单克隆抗体。

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF7, Neuro-2a, and COS-7 cell lines using Ezh2 (D2C9) XP® Rabbit mAb.

使用Ezh2 (D2C9) XP® Rabbit mAb兔单抗,免疫印迹(Western blot)分析MCF7、Neuro-2a和COS-7细胞系中Ezh2的蛋白水平。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Ezh2 (D2C9) XP® Rabbit mAb.

使用Ezh2 (D2C9) XP® Rabbit mAb兔单抗,免疫组化分析人源乳腺癌组织石蜡切片。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human cervical carcinoma using Ezh2 (D2C9) XP® Rabbit mAb.

使用Ezh2 (D2C9) XP® Rabbit mAb兔单抗,免疫组化分析人源宫颈癌组织石蜡切片。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Ezh2 (D2C9) XP® Rabbit mAb (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (blue). Actin filaments were labeled with DY-554 phalloidin (red).

使用Ezh2 (D2C9) XP® Rabbit mAb兔单抗 (绿色)和S6 Ribosomal Protein (54D2) Mouse mAb #2317 鼠单抗(蓝色)标记,共聚焦免疫荧光分析HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Ezh2 (D2C9) XP® Rabbit mAb.

使用Ezh2 (D2C9) XP® Rabbit mAb兔单抗,免疫组化分析人源淋巴瘤组织石蜡切片。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 5 µl of Ezh2 (D2C9) XP® Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,用4 x 106 NCCIT细胞的交联染色质以及5 µl Ezh2 (D2C9) XP® Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human HoxA1 Intron 1 Primers #7707、SimpleChIP® Human HoxA2 Promoter Primers #5517和SimpleChIP® Human α Satellite Repeat Primers #4486,浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号,这相当于一。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 4T1a mouse syngeneic tumor using Ezh2 (D2C9) XP® Rabbit mAb.

使用Ezh2 (D2C9) XP® Rabbit mAb兔单抗,免疫组化分析4T1a mouse syngeneic tumor组织石蜡切片。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human peripheral blood mononuclear cells untreated (left) and treated (right) with anti-human CD3 (10ug/ml, coated plates) and anti-human CD28 (5ug/ml) for 3 days at 37ºC using EZH2 (D2C9) XP® Rabbit mAb and co-stained with an anti-human CD3 antibody. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.

IF-F

IF-F

Confocal immunofluorescent analysis of mouse hippocampus (left) and cerebellum (right) using Ezh2 (D2C9) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

The polycomb group (PcG) proteins are involved in maintaining the silenced state of several developmentally regulated genes and contribute to the maintenance of cell identity, cell cycle regulation, and oncogenesis (1,2). Enhancer of zeste homolog 2 (Ezh2), a member of this large protein family, contains four conserved regions including domain I, domain II, and a cysteine-rich amino acid stretch that precedes the carboxy-terminal SET domain (3). The SET domain has been linked with histone methyltransferase (HMTase) activity. Moreover, mammalian Ezh2 is a member of a histone deacetylase complex that functions in gene silencing, acting at the level of chromatin structure (4). Ezh2 complexes methylate histone H3 at Lys9 and 27 in vitro, which is thought to be involved in targeting transcriptional regulators to specific loci (5). Ezh2 is deregulated in various tumor types, and its role, both as a primary effector and as a mediator of tumorigenesis, has become a subject of increased interest (6).

polycomb group (PcG)蛋白是涉及维持数个发育调节基因的沉默水平,并且有助于维持细胞身份(cell identity)、细胞周期调节和肿瘤发生(1,2)。zeste homolog 2 (Ezh2)的增强子是一个该蛋白家族成员,它包含四种保守区域包括domain I、domain II、一个半胱氨酸富裕的氨基酸延伸序列和羧基端SET domain (3)。SET domain已经证明与histone methyltransferase (HMTase)活性有关联。此外,哺乳类Ezh2是一个组蛋白去乙酰酶复合物的成员之一,它在基因沉默中有功能,并且在染色质结构水平起着作用(4)。在体内,Ezh2复合物使histone H3在Lys9和27位点甲基化,这被认为是涉及到调节转录因子到特异的靶位点(5)。Ezh2蛋白在不同肿瘤形式中被下调,并且具有主要效应因子和肿瘤发生的调节因子的作用已经变成一个越来越感兴趣的课题(6)。

  1. Sellers, W.R. and Loda, M. (2002) Cancer Cell 2, 349-50.
  2. Visser, H.P. et al. (2001) Br J Haematol 112, 950-8.
  3. Chen, H. et al. (1996) Genomics 38, 30-7.
  4. Tonini, T. et al. (2004) Oncogene 23, 4930-7.
  5. Müller, J. et al. (2002) Cell 111, 197-208.
  6. Kleer, C.G. et al. (2003) Proc Natl Acad Sci U S A 100, 11606-11.

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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