重组Anti-STAT1抗体[EPR4407] (ab109320)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4407] to STAT1
- Suitable for: ChIC/CUT&RUN-seq, Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-STAT1抗体[EPR4407]
参阅全部 STAT1 一抗 -
描述
兔单克隆抗体[EPR4407] to STAT1 -
宿主
Rabbit -
经测试应用
适用于: ChIC/CUT&RUN-seq, Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, A431, 293T, and MCF-7 cell lysates ICC/IF: MCF-7 cells Flow Cyt (intra): HeLa IHC-P: Human ovary carcinoma tissue. IP: MCF7. ChIC/CUT&RUN-Seq: HeLa 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.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR4407 -
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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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab109320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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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5µg |
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Flow Cyt (Intra) |
1/1000 - 1/10000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100 - 1/250.
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WB |
1/10000 - 1/50000. Predicted molecular weight: 87 kDa.
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IP |
1/10 - 1/100.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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说明 |
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ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5µg |
Flow Cyt (Intra)
1/1000 - 1/10000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/10000 - 1/50000. Predicted molecular weight: 87 kDa. |
IP
1/10 - 1/100. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
靶标
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功能
Signal transducer and activator of transcription that mediates signaling by interferons (IFNs). Following type I IFN (IFN-alpha and IFN-beta) binding to cell surface receptors, Jak kinases (TYK2 and JAK1) are activated, leading to tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize, associate with ISGF3G/IRF-9 to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of interferon stimulated genes, which drive the cell in an antiviral state. In response to type II IFN (IFN-gamma), STAT1 is tyrosine- and serine-phosphorylated. It then forms a homodimer termed IFN-gamma-activated factor (GAF), migrates into the nucleus and binds to the IFN gamma activated sequence (GAS) to drive the expression of the target genes, inducing a cellular antiviral state. -
疾病相关
Note=STAT1 deficiency results in impaired immune response leading to severe mycobacterial and viral diseases. In the case of complete deficiency, patients can die of viral disease.
Defects in STAT1 are a cause of mendelian susceptibility to mycobacterial disease (MSMD) [MIM:209950]; also known as familial disseminated atypical mycobacterial infection. This rare condition confers predisposition to illness caused by moderately virulent mycobacterial species, such as Bacillus Calmette-Guerin (BCG) vaccine and environmental non-tuberculous mycobacteria, and by the more virulent Mycobacterium tuberculosis. Other microorganisms rarely cause severe clinical disease in individuals with susceptibility to mycobacterial infections, with the exception of Salmonella which infects less than 50% of these individuals. The pathogenic mechanism underlying MSMD is the impairment of interferon-gamma mediated immunity whose severity determines the clinical outcome. Some patients die of overwhelming mycobacterial disease with lepromatous-like lesions in early childhood, whereas others develop, later in life, disseminated but curable infections with tuberculoid granulomas. MSMD is a genetically heterogeneous disease with autosomal recessive, autosomal dominant or X-linked inheritance. -
序列相似性
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
翻译后修饰
Phosphorylated on tyrosine and serine residues in response to IFN-alpha, IFN-gamma, PDGF and EGF. Phosphorylation on Tyr-701 (lacking in beta form) by JAK promotes dimerization and subsequent translocation to the nucleus. Phosphorylation on Ser-727 by several kinases including MAPK14, ERK1/2 and CAMKII on IFN-gamma stimulation, regulates STAT1 transcriptional activity. Phosphorylation on Ser-727 promotes sumoylation though increasing interaction with PIAS. Phosphorylation on Ser-727 by PKCdelta induces apoptosis in response to DNA-damaging agents.
Sumoylated by SUMO1, SUMO2 and SUMO3. Sumoylation is enhanced by IFN-gamma-induced phosphorylation on Ser-727, and by interaction with PIAS proteins. Enhances the transactivation activity.
ISGylated. -
细胞定位
Cytoplasm. Nucleus. Translocated into the nucleus in response to IFN-gamma-induced tyrosine phosphorylation and dimerization. - Information by UniProt
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数据库链接
- Entrez Gene: 6772 Human
- Omim: 600555 Human
- SwissProt: P42224 Human
- Unigene: 642990 Human
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别名
- Signal transducer and activator of transcription 1 91kD antibody
- CANDF7 antibody
- DKFZp686B04100 antibody
see all
图片
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells treated with IFN gamma (50ng/ml 1h) and 5µg of ab109320 [EPR4407]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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STAT1 was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg with 109320 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: ab109320 IP in MCF7 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab109320 in MCF7 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-STAT1 antibody [EPR4407] (ab109320) at 1/10000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : STAT1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 87 kDaLanes 1- 2: Merged signal (red and green). Green - ab109320 observed at 87 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109320 was shown to react with STAT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255346 (knockout cell lysate ab263837) was used. Wild-type HeLa and STAT1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109320 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7(Human breast adenocarcinoma cell line) cell line labeling STAT1 with ab109320 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear and cytoplasmic staining on MCF7 cells
The nuclear counterstain is DAPI (blue).
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Overlay histogram showing HeLa cells stained with ab109320 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109320, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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All lanes : Anti-STAT1 antibody [EPR4407] (ab109320) at 1/10000 dilution
Lane 1 : 293T cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 87 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted? -
ab109320 at 1/100 dilution staining STAT1 in Human ovary carcinoma by Immunohistochemistry, Paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (16)
ab109320 被引用在 16 文献中.
- Gao Y et al. Fusobacterium nucleatum stimulates cell proliferation and promotes PD-L1 expression via IFIT1-related signal in colorectal cancer. Neoplasia 35:100850 (2023). PubMed: 36371909
- Lv C et al. Narciclasine targets STAT3 via distinct mechanisms in tamoxifen-resistant breast cancer cells. Mol Ther Oncolytics 24:340-354 (2022). PubMed: 35118192
- Yan KX et al. iTRAQ-based quantitative proteomics reveals biomarkers/pathways in psoriasis that can predict the efficacy of methotrexate. J Eur Acad Dermatol Venereol 36:1784-1795 (2022). PubMed: 35666151
- Guo L et al. Epirubicin Enhances the Anti-Cancer Effects of Radioactive 125I Seeds in Hepatocellular Carcinoma via Downregulation of the JAK/STAT1 Pathway. Front Oncol 12:854023 (2022). PubMed: 35692770
- Lu T et al. Transcriptome-Based Dissection of Intracranial Aneurysms Unveils an "Immuno-Thermal" Microenvironment and Defines a Pathological Feature-Derived Gene Signature for Risk Estimation. Front Immunol 13:878195 (2022). PubMed: 35711443