Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) #13451

DNA methylation   DNMT   methl-DNA   methylated DNA  

No. Size Price
13451S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
13451 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Mouse, Endogenous 49 Rabbit IgG
IP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Specificity / Sensitivity

DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) recognizes endogenous levels of total DNMT3L protein.

DNMT3L (E1Y7Q) Rabbit mAb 兔单抗(Mouse Specific)能够检测内源性DNMT3L总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val385 of mouse DNMT3L protein.

该单克隆抗体是采用合成的与鼠源DNMT3L蛋白Val385 残基周围序列相对应的肽段免疫动物而生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from F9 and mES cells using DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific).Western blot方法检测F9和mES细胞系的提取物,使用的抗体为DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific)。

IP

IP

Immunoprecipitation of DNMT3L from F9 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) (lane 3). Lane 1 is 10% input. Western blot analysis was performed using DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific).从F9细胞提取物中免疫沉淀DNMT3L,使用的抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) 或DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) (lane 3)。Lane 1为10% input。 使用DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific)进行Western blot检测。

Western Blotting

Western Blotting

Western blot analysis of extracts from F9 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® DNMT3L siRNA I (Mouse Specific) #12307 (+), or SignalSilence® DNMT3L siRNA II (Mouse Specific) #12308 (+), using DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) confirms silencing of DNMT3L expression, while the GAPDH (D16H11) XP® Rabbit mAb is used as a loading control.Western blot方法检测经100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® DNMT3L siRNA I (Mouse Specific) #12307 (+), 或SignalSilence® DNMT3L siRNA II (Mouse Specific) #12308 (+) 转染的F9细胞系提取物,使用的抗体为DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) (上图) 或GAPDH (D16H11) XP® Rabbit mAb #5174 (下图)。使用DNMT3L (E1Y7Q) Rabbit mAb (Mouse Specific) 确认DNMT3L 表达的沉默效率, GAPDH (D16H11) XP®Rabbit mAb用作上样量对照。

Background

Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2, and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor maintenance DNA methylation.

DNMT3L is a catalytically inactive regulatory factor for the DNMT3A and DNMT3B de novo methyltransferases that is expressed at low levels in embryonic stem cells, testis, ovaries, and thymus (1,2). These de novo methyltransferases consist of a heterotetrameric complex containing two molecules of DNMT3L, and either two molecules of DNMT3A or DNMT3B (3). DNMT3L contains an amino-terminal ATRX-DNMT3-DNMT3L (ADD) domain and a carboxy-terminal methyltransferase-like domain (4-7). The methyltransferase-like domain binds to DNMT3A and DNMT3B to stimulate catalytic activity by increasing the binding of S-adenosylmethionine and DNA (4,5). The ADD domain recruits the methyltransferase complex to transcriptionally inactive regions of the genome by binding to unmethylated histone H3 Lys4 (6,7).

哺乳动物细胞中DNA胞嘧啶甲基化是一种可遗传的表观遗传修饰,并且对于基因表达、基因组印记和发育的正确调控都是至关重要的。目前已经确认了3中哺乳动物甲基转移酶家族:DNMT1、DNMT2和 DNMT3 (1,2)。DNMT1在增殖细胞中为组成型表达,是一种维持型甲基转移酶,负责为复制过程中新合成的DNA转移正确的甲基化模式。DNMT3A和DNMT3B 在胚胎干细胞中强烈表达而在成熟的体细胞组织中表达减弱。他们属于从头甲基转移酶,能够甲基化DNA上之前没有被甲基化的区域。DNMT2在成熟体细胞组织中的表达水平较低,它的失活无论是对从头DNA甲基化还是维持型DNA甲基化都没有影响。DNMT3L是DNMT3A 和 DNMT3B从头甲基转移酶的催化失活型调控因子,从而让他们在胚胎干细胞、睾丸、卵巢和胸腺中呈低水平表达(1,2)。这些从头甲基转移酶是一个异源四聚体复合物,包含两分子DNMT3L以及两分子DNMT3A或DNMT3B(3)。DNMT3L包含一个氨基末端ATRX-DNMT3-DNMT3L (ADD)结构域以及羧基端甲基转移酶样结构域(4-7)。甲基转移酶样结构域结合到DNMT3A 和 DNMT3B上通过提高S-腺苷甲硫氨酸和DNA的结合从而促进催化活性(4,5)。ADD结构域能够通过与非甲基化组蛋白H3 Lys4从而将甲基转移酶复合物募集到基因组上转录失活的区域(6,7)。

  1. Hermann, A. et al. (2004) Cell Mol Life Sci 61, 2571-87.
  2. Turek-Plewa, J. and Jagodziński, P.P. (2005) Cell Mol Biol Lett 10, 631-47.
  3. Jia, D. et al. (2007) Nature 449, 248-51.
  4. Holz-Schietinger, C. and Reich, N.O. (2010) J Biol Chem 285, 29091-100.
  5. Suetake, I. et al. (2004) J Biol Chem 279, 27816-23.
  6. Ooi, S.K. et al. (2007) Nature 448, 714-7.
  7. Otani, J. et al. (2009) EMBO Rep 10, 1235-41.

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