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HP1α Antibody

HP1α Antibody #2616

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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

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

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 (CAF-1) (7-9). In addition to contributing to heterochromatin formation and propagation, HP1 and SUV39H1 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).

Database Links

UniProt ID: P45973

Entrez-Gene Id: 23468

Limited Uses

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

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