Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Chromatin Regulation / Epigenetics

HP1β (D2F2) XP® Rabbit mAb #8676

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H M R Mk (Hm) (B) (GP) Endogenous 25 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  GP=Guinea Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

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.

Source / Purification

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

Western Blotting

Western Blotting

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

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).

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).

  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.

Application References

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

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