Revision 3

#13451Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

49

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9CWR8

Entrez-Gene Id:

54427

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

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

Species Reactivity:

Mouse

Source / Purification

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

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).

  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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalSilence is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 3
#13451

DNMT3L (E1Y7Q) Rabbit mAb

Western Blotting Image 1: DNMT3L (E1Y7Q) Rabbit mAb Expand Image
使用 DNMT3L (E1Y7Q) Rabbit mAb 对来自 F9 细胞和 mES 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 2: DNMT3L (E1Y7Q) Rabbit mAb Expand Image
使用 DNMT3L (E1Y7Q) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对经 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-)、SignalSilence® DNMT3L siRNA I #12307 (+) 或 SignalSilence® DNMT3L siRNA II #12308 (+) 转染的 F9 细胞的提取物进行蛋白质印迹分析。DNMT3L (E1Y7Q) Rabbit mAb 确认DNMT3L 表达沉默,而 GAPDH (D16H11) XP® Rabbit mAb 用作上样对照。
Immunoprecipitation Image 1: DNMT3L (E1Y7Q) Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道 2)或 DNMT3L (E1Y7Q) Rabbit mAb(泳道 3)对 F9 细胞提取物的 DNMT3L 进行免疫沉淀。泳道 1 是 10% 输入对照。使用 DNMT3L (E1Y7Q) Rabbit mAb 进行蛋白质印迹分析。