Jak/Stat Pathway
- Jak/Stat Pathway
PhosphoSitePlus®：
STAT1
- 推奨プロトコール
Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb |
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| CSTコード | 包装 | 希望納入価格 (円) |
国内在庫  2012年2月8日17時20分 現在 |
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|---|---|---|---|---|
| #9167S | 100 μL | 57,000 | ログインすると国内在庫状況がご確認いただけます。
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| #9167L | 300 μL | 137,000 |
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9167 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。 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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- 9167:
- ChIP Agarose ChIP Magnetic Flow IHC / Frozen IHC / Paraffin Immunofluorescence* Immunoprecipitation Western Blotting
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-F: 固定
IHC-P: 抗体希釈液 / 抗原賦活化
| 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)、免疫組織染色 (パラフィン) (1:400)、免疫組織染色 (凍結) (1:200)、免疫蛍光細胞染色 (IF-IC) (1:400)、フローサイトメトリー (1:200)、ChIP (1:100) |
| 種交差性 | |
|---|---|
| ヒト、マウス |
| 特異性・感度 | |
|---|---|
| 内在性レベルのTyr701 がリン酸化されたStat1 タンパク質を検出します。Tyr701 がリン酸化されたp91 Stat1 およびp84 スプライス変異も検出します。同等のチロシンがリン酸化された他のStat タンパク質とは交差しません。 |
| 検出タンパク質の分子量 | |
|---|---|
| 84 kDa、91 kDa |
| 使用抗原 | |
|---|---|
| ヒトのStat1 タンパク質のTyr701 周辺領域 (合成リン酸化ペプチド) |
| 抗体の由来 | |
|---|---|
| ウサギ |
| 貯法 | |
|---|---|
| -20℃ |
| 社内データ |
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Western Blotting
Western blot analysis of extracts from HeLa cells untreated or treated with interferon-α (IFN-α), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (upper) or Stat1 Antibody (#9172) (lower).
IHC-P (paraffin)
Immunohistochemical analysis using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb on SignalSlide (TM) Phospho-Stat1/3/5 IHC Controls #8105 (paraffin-embedded 786-0 cells untreated (left) or IFN-α treated (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma control (left) or λ phosphatase treated (right), using Phospho-Stat1 (tyr701) (58D6) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin’s lymphoma using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 in the presence of control peptide (left) or Phospho-Stat1 (Tyr701) Blocking Peptide (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded stomach (chronic gastritis), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb.
IHC-F (frozen)
Immunohistochemical analysis of frozen SKOV-3 xenograft using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of HeLa cells, untreated (blue) or IFN-α treated (green), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or IFNα-treated #9906 (right), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either 5 μl of Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb 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 human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148, 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 |
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The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.
| 使用文献 |
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- Hebel, K. et al. (2011) J Immunol 187, 5627-35. Applications: Western Blotting
本製品は試験研究用です。