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Product Usage Information
For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:2000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunoprecipitation | 1:100 |
| IHC Leica Bond | 1:100 - 1:400 |
| Immunohistochemistry (Paraffin) | 1:100 - 1:400 |
| Immunofluorescence (Immunocytochemistry) | 1:100 - 1:200 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 - 1:400 |
| Chromatin IP | 1:100 |
| Chromatin IP-seq | 1:100 |
Specificity/Sensitivity
物种反应性：
人, 小鼠, 大鼠, 猴
参考图片
Immunnohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.免疫组织化学染色分析石蜡包埋人肺癌组织,图片显示了细胞核的定位。所用抗体为Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb。
Immunohistochemical analysis of paraffin embedded human breast carcinoma, specifically endothelial cells, untreated (left) or lambda phosphatase treated (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.免疫组织化学染色分析石蜡包埋人乳腺癌组织尤其是内皮细胞，未经过处理(左图),经过lambda磷酸酶处理(右图), 所用抗体为Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb。
Western blot analysis of extracts from IFN-alpha treated Jurkat cells and HeLa cells (left), as well as EGF treated A431 cells (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb. Note that the basal phospho-Stat3 in A431 is detected by the antibody.Western免疫印迹。用Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb抗体研究IFN-alpha处理的Jurkat 细胞和 HeLa 细胞 (左图), EGF 处理 A431 细胞(右图)。注：A431 细胞中的本底phospho-Stat3 是用抗体检测。
Flow cytometric analysis of HeLa cells, untreated (blue) or IFN-α treated (green), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.流式细胞仪研究未经处理的HeLa细胞(蓝色)和经IFN-α处理的HeLa细胞(绿色)。所用抗体为Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb。
Immunohistochemical analysis of frozen H1650 xenograft using Phospho-Stat3 (Tyr705)(D3A7) XP® Rabbit mAb.免疫组织化学染色分析冷冻SKOV-3异种移植物,所用抗体为Phospho-Stat3 (Tyr705)(D3A7) XP® Rabbit mAb。
Confocal immunofluorescent analysis of HeLa cells, IFN-alpha treated (left) or untreated (right), labeled with Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (green).共聚焦免疫荧光分析经IFNalpha处理的HeLa(左图)或者未经处理的 HeLa细胞(右图) ,Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (绿色)标记。
Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine, using Phosho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.免疫组织化学染色分析石蜡包埋Apc (min/+)小鼠肠。所用抗体为Phosho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb。
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and either 10 μl of Phospho-Stat3 (Tyr705) (D3A7) XP® 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 c-Fos Promoter Primers #4663, 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.染色质免疫共沉淀。Hep G2 细胞培养至4 x 106, 并经过夜饥饿或IL-6(100 ng/ml,30 min) 处理,然后用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行免疫沉淀实验,本实验中用10 μl of Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb 或2 μl Normal Rabbit IgG #2729 抗体。用人源 IRF-1 promoter primers, SimpleChIP® Human c-Fos Promoter Primers #4663, and SimpleChIP® Human α Satellite Repeat Primers #4486对富集的DNA做real-time PCR。每个样本中沉淀的DNA量定义为相对信号与输入的总染色质相比的数值。
Immunohistochemical analysis using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb on SignalSlide® Phospho-Stat1/3/5 IHC Controls #8105 (paraffin-embedded HeLa cell pellets, untreated (left), treated with Human Interferon-α1 (hIFN-α1) #8927 (middle), or treated with Human Epidermal Growth Factor (hEGF) #8916 (right).