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#2600 Phospho-HP1γ (Ser83) Antibody

CSTコード 包装
希望納入価格 (円)
国内在庫 i
2019年8月23日15時25分 現在
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#2600S100 μL66,000
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HP1GAMMA 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性22Rabbit-20℃
種交差性 (社内試験済)
交差する可能性がある種 i

社内試験はしていませんが、配列が100%相同であるため反応すると推定される種

ヒト、マウス、ラット、サル キイロショウジョウバエ、ウシ、ウマ
2600 の推奨プロトコール i

最適な結果を得るために:Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。
推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。

注:各製品に最適化されたプロトコールをリンクしています。

 

ウェスタンブロッティング (1:1000)免疫沈降 (1:25)免疫蛍光細胞染色 (IF-IC) (1:200)

CST推奨プロトコールに欠かせない関連製品

特異性・感度
内在性レベルのSer83 (プロセシングされていないHP1γタンパク質ではSer93 に相当) がリン酸化されたHP1γタンパク質を検出します。HP1αおよび HP1βタンパク質とは交差しません。
使用抗原
ヒトのHP1γタンパク質のSer83 周辺領域のアミノ酸 (合成リン酸化ペプチド)

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社内データ

※下記の社内データは、すべて2600 の推奨プロトコールで実験した結果です。

Western Blotting

Western Blotting

Western blot analysis of whole cell extracts from HeLa cells, untreated (lanes 1 and 4), treated for 1 h with Forskolin (30 μM) and IBMX (0.5 mM) (lanes 2 and 3), or treated for 16 h with paclitaxel (500 nM) (lanes 5 and 6), using Phospho-HP1γ (Ser83) Antibody (upper panel) or HP1γ Antibody #2619 (lower panel).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left), forskolin- and IBMX-treated (center) or λ phosphatase-treated (right), using Phospho-HP1γ (Ser83) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

バックグラウンド

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).

HP1γ is phosphorylated on Ser83 by protein kinase A (PKA) in vitro, and activation of PKA by forskolin and IBMX treatment leads to increased phosphorylation in vivo (14). Phosphorylation of HP1γ on Ser83 also increases during mitosis as demonstrated by the Phospho-HP1γ (Ser83) Antibody, which shows increased immunofluorescent staining in untreated mitotic cells and increased Western blot signal in lysates from cells arrested in mitosis by treatment with paclitaxel. Phosphorylation of Ser83 only occurs on a subpopulation of HP1γ found associated with euchromatin, specifically HP1γ bound to coding regions of active genes (14). 
This phosphorylation impairs the ability of HP1γ to silence transcription and may be a marker for transcription elongation (14).

使用例
 
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Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Phospho-HP1γ (Ser83) Antibody

Metabolic Reprogramming in Disease

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