TGF-beta/Smad Signaling
| CSTコード |
包装 |
希望納入価格 (円) |
国内在庫  |
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| #9511S | 100 μL | 57,000 | |
|
| #9511L | 300 μL | 137,000 | |
9511 の推奨プロトコール
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。
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9511:
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ChIP Agarose
ChIP Magnetic
Immunoprecipitation
Western Blotting
| 用途 (希釈倍率) | |
| ウェスタンブロッティング (1:1,000)、免疫沈降 (1:100)、ChIP (1:50) |
| 種交差性 | |
| ヒト、マウス、ラット、ミンク、アフリカツメガエル |
| 特異性・感度 | |
| 内在性レベルのSer463/465 がリン酸化されたSmad1 タンパク質および同様の部位がリン酸化されたSmad5、Smad8 タンパク質を検出します。他のSmad 関連タンパク質とは交差しません。 |
| 使用抗原 | |
| ヒトのSmad5 タンパク質のSer463/465 周辺領域 (合成リン酸化ペプチド) |
Western Blotting
Western blot analysis of extracts from untransfected Xenopus cells and BMPRI/Smad1-transfected 293 cells, untreated or BMP-4-treated, using Phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad8 (Ser426/428) Antibody. (Xenopus cells provided by Dr. Malcolm Whitman, Harvard University, Massachusetts.)
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF-7 cells treated with Human BMP2 #4697 (50 ng/ml) for 1 h and either 10 μl of Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ID1 Promoter Primers #5139, human SMAD6 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.
Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β family of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate Smad1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as Smad5 and Smad8 at their corresponding sites. These phosphorylated Smads dimerize with the coactivating Smad4 and translocate to the nucleus, where they stimulate transcription of target genes (5).MAP kinases and CDKs 8 and 9 phosphorylate residues in the linker region of Smad1, including Ser206. The phosphorylation of Ser206 recruits Smurf1 to the linker region and leads to the degradation of Smad1 (6). Phosphorylation of this site also promotes Smad1 transcriptional action by recruiting YAP to the linker region (7).
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Hogan, B.L. et al. (1996) Genes Dev. 10, 1580-1594.
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Hoodless, P.A. et al. (1996) Cell 85, 489-500.
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Klemm, J.D. et al. (1998) Annu. Rev. Immunol. 16, 569-592.
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Kretzschmar, M. et al. (1997) Genes Dev. 11, 984-995.
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Whitman, M. (1998) Genes Dev. 12, 2445-2462.
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Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
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Alarcón, C. et al. (2009) Cell 139, 757-69.
PhosphoSitePlus®：
