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#54359 EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing)

 
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希望納入価格 (円)
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2019年12月13日15時35分 現在
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#54359S100 μL61,000
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#54359T20 μL39,000
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EGFR 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性175Rabbit 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 -
54359 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 (1:50)免疫蛍光細胞染色 (IF-IC) (1:500)フローサイトメトリー (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EGF Receptor タンパク質を検出します。
使用抗原
全長EGF Receptor タンパク質が発現している哺乳動物細胞

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社内データ

※下記の社内データは、すべて54359 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 A549, mIMCD-3, and CAMA-1 cells using EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-starved A549 cells that were pre-incubated (1 hr, 5 μg/ml) with EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (+), or vehicle control (-), and subsequently stimulated with hEGF #8916 (15 min, 100 ng/ml; +), using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 (upper), EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (middle), or β-actin (D6A8) Rabbit mAb #8457 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of A549 (left) and CAMA-1 (right) cells using EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of fixed, unpermeabilized (left) and live (right) CAMA-1 cells (blue), A549 cells (red), and A-431 cells (green) using EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

IF-IC

IF-IC

High-content analysis (above) of serum-starved A549 cells that were pre-incubated (1 hr) with increasing concentrations of EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) and subsequently stimulated with hEGF #8916 (15 min, 100 ng/ml). Cells were then fixed and stained for activated EGF Receptor (Phospho-EGFR Tyr1068). Data capture was performed using an ArrayScan™ VTI. Quantification of phosphorylated EGFR suggests an optimal working concentration of 5.0 μg/ml for EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) when blocking hEGF-mediated activity. Confocal immunofluorescent analysis (below) of serum-starved A549 cells that were pre-incubated (1 hr) with either 0.1 μg/ml (lower left) or 5.0 μg/ml (lower right) of EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) and subsequently stimulated with hEGF #8916 (15 min, 100 ng/ml). Cells were then fixed and stained for activated EGF Receptor (Phospho-EGFR Tyr1068) (green) and EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (red) to demonstrate the neutralizing ability of the antibody treatment. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

ELISA

ELISA

ELISA analysis of extracts from serum-starved A549 cells that were pre-incubated (1 hr, 5 μg/ml) with either vehicle control or EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (antibody-treated). The extracts were subsequently left unstimulated (red) or stimulated with hEGF #8916 (15 min, 100 ng/ml; blue), using PathScan® Phospho-EGF Receptor (panTyr) Sandwhich ELISA Kit #7911.


バックグラウンド

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

使用例
 
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DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
ArrayScan is a trademark of Thermo Fisher Scientific, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

本製品は試験研究用です。

EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing)

Metabolic Reprogramming in Disease

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