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#18338 Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb

 

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2019年3月25日15時35分 現在
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#18338S100 μL66,000
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#18338T20 μL39,000

Smad2 製品一覧

感度分子量 (kDa)抗体の由来貯法
内在性60Rabbit 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 -
18338 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 (1:100)免疫蛍光細胞染色 (IF-IC) (1:800)フローサイトメトリー (1:800)クロマチン免疫沈降 (1:50)

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Ser465 およびSer467 がリン酸化されたSMAD2 タンパク質を検出します。
使用抗原
ヒトのSMAD2 タンパク質のSer465/467 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18338 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 HaCaT cells, untreated (-) or treated with Human Transforming Growth Factor β3 (hTGF-β3) #8425 (100ng/ml, 30mins) (+), using Phospho SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb (upper) and total Smad2 (D43B4) XP® Rabbit mAb, #5339 (lower).

IP

IP

Immunoprecipitation of phospho-SMAD2 (Ser465/Ser467) from extracts of HaCaT cells treated with Human Transforming Growth Factor β1 (hTGF-β1) #8915 (10nM, 30mins). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb. Western blot analysis was performed using P-SMAD2 (Ser465/ Ser467) (E8F3R) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used for detection to avoid cross-reactivity with IgG.

IF-IC

IF-IC

Confocal immunofluorescent analysis of serum-starved HT-1080 cells, untreated (left), treated with Human Transforming Growth Factor β3 (hTGF-β3) #8425 (100 ng/mL, 20 min; center), or treated with hTGF-β3 and post-processed with λ-phosphatase (right), using Phospho-Smad2 (Ser465/467) (E8F3R) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of serum-starved HT-1080 cells, untreated (blue), treated with Human Transforming Growth Factor β3 (hTGF-β3) #8425 (100 ng/mL, 30 min; green) or pretreated with SB43152 (10 ug/mL, 30 min) and treated with hTGF-β3 (100 ng/mL, 30 min; red), using Phospho-Smad2 (Ser465/467) (E8F3R) 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® 488 Conjugate) #4412 was used as a secondary antibody.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with Human Growth Factor β1 #8915 (7 ng/ml, 1 hr) and either Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ID1 Promoter Primers #5139, Human JunB Promoter Primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

バックグラウンド

Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the receptor-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, and 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved carboxy terminal SSXS motif. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6-8).

使用例
 
関連製品
3122   Smad2 (86F7) Rabbit mAb
4486   SimpleChIP® Human α Satellite Repeat Primers
5139   SimpleChIP® Human ID1 Promoter Primers
5339   Smad2 (D43B4) XP® Rabbit mAb
7003   20X LumiGLO® Reagent and 20X Peroxide
7017   6-Tube Magnetic Separation Rack
7071   Phototope-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
8425   Human Transforming Growth Factor β3 (hTGF-β3)
9002   SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
9003   SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
9004   SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads)
9005   SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)
9006   ChIP-Grade Protein G Magnetic Beads
9007   ChIP-Grade Protein G Agarose Beads

The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation, for research use only for immunocytometry, immunohistochemistry, high content screening (HCS) analysis, or flow cytometry applications.
Tween is a registered trademark of ICI Americas, Inc.
CST 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.
SimpleChIP is a registered 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-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb

Metabolic Reprogramming in Disease

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