Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SET8 (C18B7) Rabbit mAb #2996

No. Size Price
2996S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
2996T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
2996 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 43 Rabbit IgG
IF-IC 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Bovine, Pig, Horse,

Specificity / Sensitivity

SET8 (C18B7) Rabbit mAb detects endogenous levels of total SET8 protein (both isoforms).

SET8 (C18B7) Rabbit mAb兔单抗能够检测内源性SET8总蛋白水平(两个亚型)。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to human SET8 protein. Antibodies are purified by protein A and peptide affinity chromatography.

通过合成的与人源SET8蛋白相应多肽片段去免疫动物从而制备出此单克隆抗体。通过蛋白A和多肽亲和层析纯化获得。

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT116 and RAW cells using SET8 (C18B7) Rabbit mAb.

使用SET8 (C18B7) Rabbit mAb,免疫印迹(Western blot)分析HCT116和RAW细胞中SET8 (C18B7)的蛋白水平。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with 0.5% Triton X-100 for 10 minutes prior to fixation according to Tardat et al. (2007) J. Cell Biol. 179, 1413-26 (right), using SET8 (C18B7) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

使用SET8 (C18B7) Rabbit mAb (绿色),共聚焦免疫荧光分析HeLa细胞,细胞分为未处理 (左图)或在固定之前处理0.5% Triton X-100 for 10 minutes依据Tardat et al. (2007) J. Cell Biol. 179, 1413-26 (右图)。DY-554 phalloidin标记微丝蛋白(红色)。蓝色= DRAQ5™ (DNA荧光染料)。

Background

SET domain-containing lysine methyltransferase 8 (SET8), also known as PR/SET domain-containing protein 7 (PR/SET7), is a member of a family of histone lysine methyltransferases, each of which contains a conserved catalytic SET domain originally identified in Drosophila Su[var]3-9, Enhancer of zeste, and Trithorax proteins (1-3). SET8 is a single-subunit enzyme that mono-methylates histone H4 on Lys20, preferably on nucleosomal substrates (1-3). SET8 protein levels and Histone H4 Lys20 methylation are cell cycle regulated, both increasing in S phase and peaking at G2/M phase (4,5). SET8 interacts with the PCNA protein, associates with sites of active DNA synthesis and is required for DNA replication and genome stability during S phase (5-7). Inhibition of SET8 using shRNA results in arrest of replication forks, induction of double-stranded DNA breaks and a Chk1-mediated cell-cycle arrest in S and G2/M phases of the cell cycle (6,7). Furthermore, SET8 methylates p53 on Lys382, down regulating the pro-apoptotic and checkpoint activation functions of p53 (8). In response to DNA damage, SET8 expression levels decrease, allowing p53 to activate checkpoints and/or apoptosis (8). Both the methylation of histone H4 Lys20 and p53 appear to be important for the functions of SET8 in S phase.

SET domain-containing lysine methyltransferase 8 (SET8)也称为PR/SET domain-containing protein 7 (PR/SET7),它是一个组蛋白赖氨酸甲基转移酶家族成员,它们中每一个都包含一个保守催化的SET结构,该结构起初被鉴定在Drosophila Su[var]3-9、Enhancer of zeste和Trithorax蛋白中(1-3)。SET8是一个单亚单位激酶,它能使histone H4蛋白Lys20位点发生单甲基化,更适应在核小体结构底物上(1-3)。SET8蛋白水平和Histone H4蛋白Lys20位点甲基化是被细胞周期调节的,在S期处于增长期而到G2/M期达到最高峰(4,5)。SET8与PCNA蛋白相互作用,这与活化的DNA合成的位点有关联,并且对于在S期中DNA复制和基因组稳定性是需要的(5-7)。使用shRNA进行SET8的抑制可导致复制叉的阻止、双链DNA诱导的中断和在细胞周期的S和G2/M期中一个Chk1介导的细胞周期停滞 (6,7)。更进一步,SET8可使p53蛋白在Lys382位点甲基化,这可下调p53蛋白促凋亡和监测点激活功能(8)。在DNA损伤反应下,SET8表达水平下降,这允许p53去激活检测点和/或凋亡(8)。histone H4蛋白Lys20位点的甲基化和p53出现对于在S期中SET8的功能是非常重要的。

  1. Fang, J. et al. (2002) Curr Biol 12, 1086-99.
  2. Xiao, B. et al. (2005) Genes Dev 19, 1444-54.
  3. Couture, J.F. et al. (2005) Genes Dev 19, 1455-65.
  4. Rice, J.C. et al. (2002) Genes Dev 16, 2225-30.
  5. Huen, M.S. et al. (2008) J Biol Chem 283, 11073-7.
  6. Tardat, M. et al. (2007) J Cell Biol 179, 1413-26.
  7. Jørgensen, S. et al. (2007) J Cell Biol 179, 1337-45.
  8. Shi, X. et al. (2007) Mol Cell 27, 636-46.

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

U.S. Patent No. 5,675,063.

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

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