| 货号 | 8676T |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human,Mouse,Rat,Monkey, |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IP , IF-IC , ChIP , ChIP-seq |
| 目标/特异性 | 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. |
| 使用方法 | WB(1:1000) IP (1:50) IF-IC (1:800) ChIP (1:50) ChIP-seq (1:50) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | 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). |
| 运输条件 | 1 |
| 存放说明 | -20C |
| 计算分子量 | 25 |
| 参考文献 | 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, 120-123. 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, 1132-6. 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. |
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 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 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. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hela cells and either 10 µl of HP1β (D2F2) XP® Rabbit mAb or 10 μl of Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb #13969, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for HP1β ChIP-seq and 50ng enriched ChIP DNA for H3K9me3 ChIP-seq using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. HP1β and H3K9me3 are known to associate with each other on chromatin. The figure shows binding of both HP1β and H3K9me3 across ZNF genes, known target genes of both HP1β and H3K9me3. For additional ChIP-seq tracks, please download the product data sheet. |
