Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

HP1β (D2F2) XP® Rabbit mAb #8676

No. Size Price
8676S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
8676 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 25 Rabbit IgG
IP 1:50
IF-IC 1:800
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

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

Specificity / Sensitivity

HP1β (D2F2) XP® Rabbit mAb recognizes endogenous levels of total HP1β protein. This antibody does not cross-react with other HP1 proteins, including HP1α and HP1γ.

HP1β (D2F2) XP® Rabbit mAb兔单抗能够检测内源性HP1β总蛋白水平。该抗体不与其它HP1蛋白发生交叉反应,包括HP1α和HP1γ。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln69 of human HP1β protein.

该单克隆抗体是采用合成的与人源HP1β蛋白Gln69残基周围序列相对应的肽段免疫动物而生产的。

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells using HP1β (D2F2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

使用HP1β (D2F2) XP® Rabbit mAb兔单抗 (绿色),共聚焦免疫荧光分析C2C12细胞。DY-554 phalloidin标记微丝蛋白(红色)。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using HP1β (D2F2) XP® Rabbit mAb.

使用HP1β (D2F2) XP® Rabbit mAb兔单抗,免疫印迹(Western blot)分析不同细胞中HP1β (D2F2)的蛋白水平。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 mES cells and either 10 µl of HP1β (D2F2) XP® Rabbit mAb #8676 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® Mouse MEST Intron 1 Primers #12870, SimpleChIP® Mouse MEST Promoter Primers #12928, mouse Intracisternal A-Particle (IAP) LTR promoter primers, and mouse primers specific to a non-genic, non-repetitive region. 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.

Background

Heterochromatin protein 1 (HP1) is a family of heterochromatic adaptor molecules involved in both gene silencing and higher order chromatin structure (1). All three HP1 family members (α, β, and γ) are primarily associated with centromeric heterochromatin; however, HP1β and γ also localize to euchromatic sites in the genome (2,3). HP1 proteins are approximately 25 kDa in size and contain a conserved amino-terminal chromodomain, followed by a variable hinge region and a conserved carboxy-terminal chromoshadow domain. The chromodomain facilitates binding to histone H3 tri-methylated at Lys9, a histone "mark" closely associated with centromeric heterochromatin (4,5). The variable hinge region binds both RNA and DNA in a sequence-independent manner (6). The chromoshadow domain mediates the dimerization of HP1 proteins, in addition to binding multiple proteins implicated in gene silencing and heterochromatin formation, including the SUV39H histone methyltransferase, the DNMT1 and DNMT3a DNA methyltransferases, and the p150 subunit of chromatin-assembly factor-1 (CAF1) (7-9). In addition to contributing to heterochromatin formation and propagation, HP1 and SUV39H are also found complexed with retinoblastoma (Rb) and E2F6 proteins, both of which function to repress euchromatic gene transcription in quiescent cells (10,11). HP1 proteins are subject to multiple types of post-translational modifications, including phosphorylation, acetylation, methylation, ubiquitination, and sumoylation, suggesting multiple means of regulation (12-14).

Heterochromatin protein 1 (HP1)作为一类异染色质调节因子参与了基因的沉默和染色质高级结构(1)。HP1家族总共有三个成员(α, β和γ),它们主要和异染色质的着丝粒相结合;但是HP1β和γ也在基因组的常染色质中存在(2,3)。HP1蛋白大约有25KD,并且都有保守的氨基酸末端染色质结构域,后面跟了一个可变的铰链区和一个保守的碳末端chromoshadow结构域。染色质结构域可以帮助结合到在Lys9位点发生了3甲基化的H3上,这是一个和着丝粒异染色质密切相关的组蛋白“标志”(4,5)。这个可变的铰链区可以通过sequence-independent的方式与RNA、DNA结合(6)。Chromoshadow 结构域除了结合包括SUV39 Hhistone methyltransferase、DNMT1、DNMT3a DNA methyltransferases 以及 the p150 subunit of chromatin-assembly factor-1 (CAF1) 在内的多种与基因沉默和异染色质形成的蛋白外,还介导HP1蛋白的二聚体化(7-9)。除了参与异染色质的形成和扩展,HP1和SUV39H同样也发现和retinoblastoma (Rb)与 E2F6 proteins这两种在静止期细胞中抑制基因转录的蛋白共同形成复合物(10,11)。HP1蛋白受到多种翻译后修饰包括:磷酸化,乙酰化,甲基化,泛素化和苏木化,这表明有多重调节方式(12-14)。

  1. Maison, C. and Almouzni, G. (2004) Nat. Rev. Mol. Cell Biol. 5, 296-304.
  2. Minc, E. et al. (2000) Cytogenet. Cell Genet. 90, 279-284.
  3. Nielsen, A.L. et al. (2001) Mol. Cell 7, 729-739.
  4. Lachner, M. et al. (2001) Nature 410, 116-120.
  5. Bannister, A.J. et al. (2001) Nature 410, 120-124.
  6. Muchardt, C. et al. (2002) EMBO Rep. 3, 975-981.
  7. Yamamoto, K. and Sonoda, M. (2003) Biochem. Biophys. Res. Commun. 301, 287-292.
  8. Fuks, F. et al. (2003) Nucleic Acids Res. 31, 2305-2312.
  9. Murzina, N. et al. (1999) Mol. Cell 4, 529-540.
  10. Nielsen, S.J. et al. (2001) Nature 412, 561-565.
  11. Ogawa, H. et al. (2002) Science 296, 1132-1136.
  12. Minc, E. et al. (1999) Chromosoma 108, 220-234.
  13. Zhao, T. et al. (2001) J. Biol. Chem. 276, 9512-9518.
  14. Lomberk, G. et al. (2006) Nat. Cell Biol. 8, 407-415.

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

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