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#16177 Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb

 

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2019年3月20日11時40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
16177 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)免疫組織染色 (パラフィン) (1:320)免疫蛍光細胞染色 (IF-IC) (1:800)フローサイトメトリー (1:800)

下記ステップについては、データシートの右側もあわせてご参照ください。

IHC-P: 抗体希釈液 / 抗原賦活化

CST推奨プロトコールに欠かせない関連製品

特異性・感度
Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb recognizes endogenous levels of SQSTM1/p62 protein only when phosphorylated at Ser349. Staining of mitotic cells is observed by immunohistochemstry. The specificity of this staining is unknown.
使用抗原
Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser349 of human SQSTM1/62 protein.

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社内データ

※下記の社内データは、すべて16177 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 wild-type MEF and MEF SQSTM1/p62 KO cell lines, untreated (-) or treated with sodium arsenite (15 μM, 18 hr; +), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb (upper), SQSTM1/p62 (D1Q5S) Rabbit mAb #39749 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). MEF SQSTM1/p62 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston MA.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC-3 cells treated with sodium arsenite (15 μM, indicated times) using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb (upper), 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 PC-3 cells treated with sodium arsenite (15 μM, 18 hr) with lysates that were untreated (-) or treated with lambda-phosphatase and calf intestinal phosphatase (λPPase/CIP; +) using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb (upper), SQSTM1/p62 (D5L7G) Mouse mAb #88588 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untreated (left, negative) or treated with sodium arsenite (right, positive), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of PC-3 cells, untreated (left), treated with sodium arsenite (15 μM, 18 hr; center), or treated with sodium arsenite and post-processed with λ-phosphatase (right), using Phospho-SQSTM1 (Ser349) (E7M1A) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 650 Phalloidin #12956 (red). Blue = DAPI #4083 (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of PC-3 cells, untreated (blue) or treated with sodium arsenite (15 µM, 18 hr; green), using Phospho-SQSTM1 (Ser349) (E7I4Z) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

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 Ser349 (Ser351 in mouse) during oxidative stress increases its binding to KEAP1, thereby increasing NRF2 activity (13).

使用例
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13079   Immunohistochemistry Application Solutions Kit (Rabbit)
13121   Phospho-SQSTM1/p62 (Thr269/Ser272) Antibody
14166   Hematoxylin
14177   SignalStain® Mounting Medium
14746   SignalStain® Citrate Unmasking Solution (10X)
15019   Animal-Free Blocking Solution (5X)
23214   SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific)
39749   SQSTM1/p62 (D1Q5S) Rabbit mAb
39786   Phospho-SQSTM1/p62 (Ser403) (D8D6T) Rabbit mAb
5114   SQSTM1/p62 Antibody
6394   SignalSilence® SQSTM1/p62 siRNA I
6399   SignalSilence® SQSTM1/p62 siRNA II
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7695   SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred)
7727   Biotinylated Protein Ladder Detection Pack
8025   SQSTM1/p62 (D5E2) Rabbit mAb
8059   SignalStain® DAB Substrate Kit
8112   SignalStain® Antibody Diluent
8114   SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
88588   SQSTM1/p62 (D5L7G) Mouse mAb
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)

Tween is a registered trademark of ICI Americas, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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 (Ser349) (E7M1A) Rabbit mAb

Immune Cell Signaling Pathways

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