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#39786 Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb

 

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2019年3月22日15時35分 現在
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SQSTM1 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性62Rabbit IgG-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 -
39786 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)免疫蛍光細胞染色 (IF-IC) (1:400)

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Ser403 がリン酸化されたSQSTM1/p62 タンパク質を検出します。
使用抗原
ヒトのSQSTM1/p62 タンパク質のSer403 周辺領域 (合成ペプチド)

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社内データ

※下記の社内データは、すべて39786 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 extracts from 293T cells, mock transfected (-) or transfected with constructs expressing GFP-tagged human SQSTM1/p62 (hSQSTM1/p62-GFP; +) and human GST-tagged IKK-ε (hIKK-ε-GST; +) as indicated, using Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C2C12, and H-4-II-E cells, untreated (-) or treated with Chloroquine #14774 (50 μM, overnight; +), using Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb (upper), total SQSTM1/p62 (D1Q5S) Rabbit mAb #39749 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of STF-66247-treated 786-O cell extracts, that were either untreated (-) or treated with Lambda Phosphatase plus Calf Intestinal Phosphatse ( λPPase/CIP; +), using Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb (upper), total SQSTM1/p62 (D5L7G) Mouse mAb #88588 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from 786-O cells, untreated (-) or treated with STF-66247 (1 μM, indicated times), using Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb (upper), total SQSTM1 (D5L7G) Mouse mAb #88588 (middle), or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of 786-O cells, untreated (left) or treated with STF-66247 (1 μM, 18 hr; right), using Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

バックグラウンド

Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein involved in cell signaling, oxidative stress, and autophagy (1-4). It was first identified as a protein that binds to the SH2 domain of p56Lck (5) and independently found to interact with PKCζ (6,7). SQSTM1 was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins and triggering degradation of proteins through the proteasome or lysosome (8). Interaction between SQSTM1 and TRAF6 leads to the K63-linked polyubiquitination of TRAF6 and subsequent activation of the NF-κB pathway (9). Protein aggregates formed by SQSTM1 can be degraded by the autophagosome (4,10,11). SQSTM1 binds autophagosomal membrane protein LC3/Atg8, bringing SQSTM1-containing protein aggregates to the autophagosome (12). Lysosomal degradation of autophagosomes leads to a decrease in SQSTM1 levels during autophagy; conversely, autophagy inhibitors stabilize SQSTM1 levels. Studies have demonstrated a link between SQSTM1 and oxidative stress. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (3). Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity.

Phosphorylation of SQSTM1 at Ser403 increases its affinity for polyubquitinated chains, resulting in enhanced autophagic clearance (13,14). This site has been reported to be phosphorylated by casein kinase 2 (CK2), as well as by the innate immunity regulator TBK-1.

使用例
 
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88588   SQSTM1/p62 (D5L7G) Mouse mAb
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DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

本製品は試験研究用です。

Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb

Metabolic Reprogramming in Disease

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