Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

HP1α Antibody #2616

Antigen p25   CBX5   Heterochromatin protein 1 homolog alpha   HP1A  

No. Size Price
2616S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
2616 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 25 Rabbit
IP 1:25
IHC-P 1:200
F 1:100
IF-IC 1:200
ChIP 1:25

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

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

Specificity / Sensitivity

HP1 alpha antibody detects endogenous levels of total HP1alpha protein. The antibody does not cross-react with HP1 beta or HP1 gamma proteins.

HP1 alpha antibody检测内源性HP1alpha总蛋白水平。该抗体不能与HP1 beta或HP1 gamma蛋白发生交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of human HP1alpha. Antibodies are purified by protein A and peptide affinity chromatography.

通过合成的与人源HP1alpha蛋白羧基末端周围相应的多肽片段去免疫动物从而制备出多克隆抗体。通过蛋白A和多肽亲和层析纯化获得。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated HeLa cells, using HP1alpha antibody (blue) compared to a nonspecific negative control antibody (red).

与非特异阴性control antibody (红色)比较,使用HP1alpha antibody (blue)标记,流式细胞仪分析非处理的HeLa细胞。

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa, NIH/3T3, C6 and COS cells, using HP1alpha antibody.

使用HP1alpha antibody,免疫印迹(Western blot)分析HeLa, NIH/3T3, C6和COS细胞中HP1alpha蛋白水平。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using HP1alpha Antibody in the presence of control peptide (left) or HP1 alpha blocking peptide #1004 (right).

使用HP1alpha Antibody,其分别与control peptide (左图)或HP1 alpha blocking peptide #1004 (右图)孵育,免疫组化分析人源乳腺癌组织石蜡切片。

IF-IC

IF-IC

Confocal immunofluorescent image of HeLa cells labeled with HP1α antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

使用HP1α antibody (绿色)标记,共聚焦免疫荧光分析HeLa细胞。Alexa Fluor® 555 phalloidin标记微丝蛋白(红色)。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 mES cells and either 20 µl of HP1α Antibody #2616 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 each contains 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 on 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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