重组Anti-STAT3 (phospho Y705)抗体[EP2147Y] (ab76315)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2147Y] to STAT3 (phospho Y705)
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, Dot blot, ICC/IF, ELISA
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-STAT3 (phospho Y705)抗体[EP2147Y]
参阅全部 STAT3 一抗 -
描述
兔单克隆抗体[EP2147Y] to STAT3 (phospho Y705) -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IP, IHC-P, Dot blot, ICC/IF, ELISAmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab179551) -
阳性对照
- WB: HeLa cell lysate treated with alpha-interferon. IHC-P: Rat colon tissue, mouse colon tissue, mouse spleen tissue and human thyroid carcinoma tissue. ICC/IF: HeLa cells treated with alpha-interferon; U251 cells. IP: A431 cells treated with EGF. Flow Cyt (intra): A431 cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP2147Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab76315于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/500.
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WB | (9) |
1/2000 - 1/20000. Predicted molecular weight: 88 kDa.Can be blocked with STAT3 (phospho Y705) peptide (ab179551).
Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls. |
IP |
1/20.
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IHC-P | (6) |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Dot blot |
1/1000.
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ICC/IF | (2) |
1/500.
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ELISA | (1) |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
1/500. |
WB
1/2000 - 1/20000. Predicted molecular weight: 88 kDa.Can be blocked with STAT3 (phospho Y705) peptide (ab179551). Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls. |
IP
1/20. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Dot blot
1/1000. |
ICC/IF
1/500. |
ELISA
Use at an assay dependent concentration. |
靶标
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功能
Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. -
组织特异性
Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. -
疾病相关
Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
Autoimmune disease, multisystem, infantile-onset -
序列相似性
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
翻译后修饰
Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling. -
细胞定位
Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1. - Information by UniProt
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数据库链接
- Entrez Gene: 6774 Human
- Entrez Gene: 20848 Mouse
- Entrez Gene: 25125 Rat
- Omim: 102582 Human
- SwissProt: P40763 Human
- SwissProt: P42227 Mouse
- SwissProt: P52631 Rat
- Unigene: 463059 Human
see all -
别名
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
see all
图片
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All lanes : Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315) at 1/20000 dilution (purified)
Lane 1 : Un-treated HeLa (Human epithelial cell line from cervix adenocarcinoma )cell lysate
Lane 2 : HeLa cell lysate - treated with IFN-a
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 88 kDa
Observed band size: 88 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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NEAT1 influences STAT3 binding to endocytosis-related genes. The siNEAT1 and siCTRL cells were collected for ChIP assays to analyse the relative fold enrichment of the CAV2 (a), TGFB2 (b), or TGFBR1 promoter (c) with anti-STAT3 or RNAP II antibodies. The data points represent the mean values determined from three independent experiments. The data are presented as the mean ± SD. d The U251 cells were fixed and incubated with anti-STAT3 (red), anti-H3K27Ac (green) or anti-H3K27Cro antibodies (green) before the confocal analysis. The intensity plots for the red and green channels were analysed with the ImageJ software. Scale bars 10 μm. e After transfection with the siNEAT1v2 or negative control siRNA, the U251 cells were fixed and incubated with anti-STAT3 (red) or anti-H3K27Ac antibodies (green) before the confocal analysis. The intensity plots for the red and green channels were analysed with the ImageJ software. Scale bars 10 μm. f After 24 h of incubation with 80 µm crotonyl-CoA or the mock control, the U251 cell lysates were harvested and subjected to an immunoprecipitation assay with anti-STAT3 or anti-IgG antibodies. The retrieval of STAT3, H3K27Ac, H3K27Cro+ and Histone H3 by endogenous STAT3 and IgG was measured by western blotting. *p < 0.001
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Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) +/- IFN-α (50 ng/mL, 5 minutes) cells labelling STAT3 (phospho Y705) with ab76315 at 1/500 (4.3 μg/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1500) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded mouse colon tissue stained for STAT3 using ab76315 at 1/1000 dilution followed by a Horse radish peroxidase conjugated Goat anti-Rabbit secondary antibody at 1/400 dilution.
Antigen Retrieval: Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA 70C 2hr.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labeling STAT3 (phospho Y705) with ab76315 at 1/100 (nuclear staining). Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody. Counterstained with hematoxylin.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/mL EGF for 10min (Red) / Untreated control (Green) cells labelling STAT3 with ab76315 at 1/500 dilution (0.1µg) (Red) and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Dot blot analysis of STAT3 single phospho peptide pY705 (lane 1) and STAT3 non-phospho peptide (lane 2) with ab76315 at 1/1000. Blocking and dilution buffer was 5% NFDM/TBST. The secondary antibody used was ab97051 peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/100,000.
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Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue using untreated (left) or alkaline phosphatase-treated (right) labeling STAT3 (phospho Y705) with ab76315 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H& L (HRP) (ab97051) at 1/500 dilution. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.
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ab76315 (purified) at 1/30 immunoprecipitating STAT3 (phospho Y705) in A431 (Human epidermoid carcinoma cell line) cell lysate treated with EGF. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).
Blocking/Dilution buffer: 5% NFDM/TBST.
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Tissue Microarrays stained for "Anti-STAT3 (phospho Y705) antibody [EP2147Y]” using "ab76315" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab97051 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab76315 at +4°C overnight followed by Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (535)
ab76315 被引用在 535 文献中.
- Li X et al. Mechanistic Insights of Qingre Jiedu Recipe Based on Network Pharmacology Approach against Heart Failure. Evid Based Complement Alternat Med 2022:9024394 (2022). PubMed: 35140803
- Zhang Z et al. Rifaximin protects SH-SY5Y neuronal cells from iron overload-induced cytotoxicity via inhibiting STAT3/NF-κB signaling. Cell Biol Int 46:1062-1073 (2022). PubMed: 35143099
- Li Y et al. microRNA-378a-3p regulates the progression of hepatocellular carcinoma by regulating PD-L1 and STAT3. Bioengineered 13:4730-4743 (2022). PubMed: 35184646
- Chen Q et al. Traditional Chinese Medicine Pien-Tze-Huang Inhibits Colorectal Cancer Growth and Immune Evasion by Reducing β-catenin Transcriptional Activity and PD-L1 Expression. Front Pharmacol 13:828440 (2022). PubMed: 35185580
- Chen Q et al. Mechanism of Synsepalum dulcificum Daniell. Inhibiting Lung Adenocarcinoma. Evid Based Complement Alternat Med 2022:5242179 (2022). PubMed: 35190747