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#92340 Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb

 

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2019年3月22日15時35分 現在
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Atg14 製品一覧 | #92340 が入っているAntibody Sampler キット一覧

感度分子量 (kDa)抗体の由来貯法
内在性65Rabbit 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

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92340 の推奨プロトコール i

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ウェスタンブロッティング (1:1000)免疫蛍光細胞染色 (IF-IC) (1:800)

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特異性・感度
Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb recognizes endogenous levels of Atg14 protein only when phosphorylated at Ser29.
使用抗原
Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser29 of human Atg14 protein.

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社内データ

※下記の社内データは、すべて92340 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 Atg14 protein (hAtg14-GFP; +) or mouse ULK1 protein (mULK1; +), using Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb (upper), Atg14 (D1A1N) Rabbit mAb (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT 116 and HCT 116/Atg14 shRNA knockout cells, untreated (-) or starved using Earles Basic Salt Solution (EBSS, 2 hr; +) and the ULK1 inhibitor SBI-020695 #29089 (50 μM, 2 hr; +) as indicated, using Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb (upper), Atg14 (D1A1N) Rabbit mAb #96752 (middle), or β-Actin (D6A8) Rabbit mAb (lower). HCT 116/Atg14 shRNA knockout cells were kindly provided by Dr. Do-Hyung Kim, University of Minnesota, Minneapolis, MN.

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT 116 cells, untreated (-) or treated with lambda-phosphatase and calf intestinal phosphatase (λ-phosphatase/CIP; +), using Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb (upper), Atg14 (D1A1N) Rabbit mAb #96752 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from Saos-2 cells, untreated (-) or starved using Earles Basic Salt Solution (EBSS, 2 hr), using Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb (upper), Atg14 (D1A1N) Rabbit mAb #96752 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HCT 116 Atg14 wild-type cells, untreated (left, low-expressing) or treated with Torin 1 #14379 (250 nM, 2 hr; middle-left, high-expressing), HCT 116/Atg14 shRNA knockout cells treated with Torin 1 (middle-right, negative), or HCT 116 Atg14 wild-type cells post-processed with λ-phosphatase (2 hr; right, negative), using Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue). HCT 116/Atg14 shRNA knockout cells were kindly provided by Dr. Do-Hyung Kim, University of Minnesota, Minneapolis, MN.

バックグラウンド

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but is also associated with a number of physiological processes including development, differentiation, neurodegeneration, infection and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and is directed by a number of autophagy-related (Atg) genes. These proteins are involved in the formation of autophagosomes, cytoplasmic vacuoles that are delivered to lysosomes for degradation. The class III type phosphoinositide 3-kinase (PI3K) Vps34 regulates vacuolar trafficking and autophagy (4,5). Multiple proteins associate with Vsp34, including p105/Vsp15, Beclin-1, UVRAG, Atg14, and Rubicon, to determine Vsp34 function (6-12). Atg14 and Rubicon were identified based on their ability to bind to Beclin-1 and participate in unique complexes with opposing functions (9-12). Rubicon, which localizes to the endosome and lysosome, inhibits Vps34 lipid kinase activity; knockdown of Rubicon enhances autophagy and endocytic trafficking (11,12). In contrast, Atg14 localizes to autophagosomes, isolation membranes and ER, and can enhance Vps34 activity. Knockdown of Atg14 inhibits starvation-induced autophagy (11,12).

The serine/threonine kinase ULK1 phosphorylates Atg14 at Ser29 to promote autophagsome formation (13).

使用例
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13155   Phospho-Atg14 (Ser29) Antibody
5504   Atg14 Antibody
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
96752   Atg14 (D1A1N) Rabbit mAb

Tween is a registered trademark of ICI Americas, Inc.
ProLong 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-Atg14 (Ser29) (D4B8M) Rabbit mAb

Metabolic Reprogramming in Disease

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