HP1α Antibody #2616
- WB
- IP
- IHC
- IF
- F
- ChIP
Supporting Data
| REACTIVITY | H M R Mk |
| SENSITIVITY | Endogenous |
| MW (kDa) | 25 |
| SOURCE | Rabbit |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IHC-Immunohistochemistry
- IF-Immunofluorescence
- F-Flow Cytometry
- ChIP-Chromatin Immunoprecipitation
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Mk-Monkey
Product Information
Product Usage Information
For optimal ChIP results, use 20 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:25 |
| Immunohistochemistry (Paraffin) | 1:200 |
| Immunofluorescence (Immunocytochemistry) | 1:200 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 |
| Chromatin IP | 1:25 |
Storage
Protocol
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.
Species Reactivity:
Human, Mouse, Rat, Monkey
The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.
Species predicted to react based on 100% sequence homology:
Bovine
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.
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).
- Maison, C. and Almouzni, G. (2004) Nat. Rev. Mol. Cell Biol. 5, 296-304.
- Minc, E. et al. (2000) Cytogenet. Cell Genet. 90, 279-284.
- Nielsen, A.L. et al. (2001) Mol. Cell 7, 729-739.
- Lachner, M. et al. (2001) Nature 410, 116-120.
- Bannister, A.J. et al. (2001) Nature 410, 120-124.
- Muchardt, C. et al. (2002) EMBO Rep. 3, 975-981.
- Yamamoto, K. and Sonoda, M. (2003) Biochem. Biophys. Res. Commun. 301, 287-292.
- Fuks, F. et al. (2003) Nucleic Acids Res. 31, 2305-2312.
- Murzina, N. et al. (1999) Mol. Cell 4, 529-540.
- Nielsen, S.J. et al. (2001) Nature 412, 561-565.
- Ogawa, H. et al. (2002) Science 296, 1132-1136.
- Minc, E. et al. (1999) Chromosoma 108, 220-234.
- Zhao, T. et al. (2001) J. Biol. Chem. 276, 9512-9518.
- Lomberk, G. et al. (2006) Nat. Cell Biol. 8, 407-415.
Limited Uses
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