Name: Anti-STAT1 (phospho S727) antibody [EPR3146]
Brand: RabMAb
SKU: ab109461
Rating: 4 (1 reviews)

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ChIC/CUT&RUN sequencing - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR3146] to STAT1 (phospho S727)
Suitable for: WB, IHC-P, ChIC/CUT&RUN-seq, Dot blot
Reacts with: Mouse, Rat, Human

Carrier Free

产品名称

Anti-STAT1 (phospho S727)抗体[EPR3146]
参阅全部 STAT1 一抗
描述

兔单克隆抗体[EPR3146] to STAT1 (phospho S727)
宿主

Rabbit
特异性

A phospho specific peptide corresponding to residues surrounding Serine 727 of human Stat-1 was used as an immunogen. This antibody only detects Stat-1 phosphorylated at Serine 727.
经测试应用

适用于: WB, IHC-P, ChIC/CUT&RUN-seq, Dot blotmore details
不适用于: ICC/IF
种属反应性

与反应: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: HeLa cell lysate. Rat and mouse brain lysate. IHC-P: Rat and mouse colon tissue. Human breast carcinoma and stomach adenocarcinoma tissue. ChIC/CUT&RUN-Seq: HeLa cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR3146
同种型

IgG
研究领域

Alternative Versions
- Anti-STAT1 (phospho S727) antibody [EPR3146] - BSA and Azide free (ab215820)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Abpromise™承诺保证使用ab109461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
WB	(1)	1/1000 - 1/10000. Detects a band of approximately 91 kDa (predicted molecular weight: 87 kDa).
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat up to 98 °C, below boiling, and then let cool for 10-20 minutes.
ChIC/CUT&RUN-seq		Use at an assay dependent concentration. 5µg
Dot blot		Use at an assay dependent concentration.

说明
WB 1/1000 - 1/10000. Detects a band of approximately 91 kDa (predicted molecular weight: 87 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat up to 98 °C, below boiling, and then let cool for 10-20 minutes.
ChIC/CUT&RUN-seq Use at an assay dependent concentration. 5µg
Dot blot Use at an assay dependent concentration.

应用说明

Is unsuitable for ICC/IF.

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.
Target information above from: UniProt accession P42224 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 6772 Human
- Entrez Gene: 20846 Mouse
- Entrez Gene: 25124 Rat
- Omim: 600555 Human
- SwissProt: P42224 Human
- SwissProt: P42225 Mouse
- Unigene: 642990 Human
别名
- Signal transducer and activator of transcription 1 91kD antibody
- CANDF7 antibody
- DKFZp686B04100 antibody
- IMD31A antibody
- IMD31B antibody
- IMD31C antibody
- ISGF 3 antibody
- ISGF-3 antibody
- OTTHUMP00000163552 antibody
- OTTHUMP00000165046 antibody
- OTTHUMP00000165047 antibody
- OTTHUMP00000205845 antibody
- Signal transducer and activator of transcription 1 91kDa antibody
- Signal transducer and activator of transcription 1 antibody
- Signal transducer and activator of transcription 1, 91kD antibody
- Signal transducer and activator of transcription 1-alpha/beta antibody
- STAT 1 antibody
- Stat1 antibody
- STAT1_HUMAN antibody
- STAT91 antibody
- Transcription factor ISGF 3 components p91 p84 antibody
- Transcription factor ISGF-3 components p91/p84 antibody
- Transcription factor ISGF3 components p91/p84 antibody
- XStat1 antibody
see all

ChIC/CUT&RUN sequencing - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

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) treated with IFN gamma (50ng/ml 1h) cells and 5µg of ab109461 [EPR3146]. 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.
Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

All lanes : Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/1000 dilution

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate (treated with Alkaline Phosphatase for 1 hour)
Lane 3 : Mouse brain lysate
Lane 4 : Mouse brain lysate (treated with Alkaline Phosphatase for 1 hour)

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 87 kDa
Observed band size: 91 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST.
Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

All lanes : Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/1000 dilution

Lane 1 : Rat brain lysate
Lane 2 : Rat brain lysate (treated with Alkaline Phosphatase for 1 hour)

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 87 kDa
Observed band size: 91 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST.
Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

All lanes : Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/5000 dilution (purified)

Lane 1 : Untreated HeLa whole cell lysate
Lane 2 : HeLa whole cell lysate treated with etoposide
Lane 3 : HeLa whole cell lysate treated with etoposide, followed by membrane treatment with phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 87 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

Blocking buffer: 2% BSA/TBST
Dilution buffer: 2% BSA/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Immunohistochemical staining of paraffin embedded rat colon with purified ab109461 at a working dilution of 1/200. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Immunohistochemical staining of paraffin embedded mouse colon with purified ab109461 at a working dilution of 1/200. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab109461 at a working dilution of 1/200. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

All lanes : Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/10000 dilution (unpurified)

Lane 1 : Untreated HeLa (human cervix adenocarcinoma)
Lane 2 : HeLa (human cervix adenocarcinoma) membrane treated with Alkaline Phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1500 dilution

Developed using the ECL technique.

Predicted band size: 87 kDa
Observed band size: 91 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

The lower section shows STAT1 detected with ab92506, anti-STAT1 antibody, to confirm that the same amount of lysate is used in each lane.
Blocking and dilution biffer: 5% NFDM/TBST.
Dot Blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Dot Blot analysis of Lane 1: STAT1 (pS727) phospho peptide and Lane 2: STAT1 non-phospho peptide, labeling STAT1 (phospho S727) with ab109461 at 1/1000 dilution. 5% NFDM/TBST was used as the blocking and diluting buffer. ab97051, a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody was used at 1/100000 dilution. Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Unpurified ab109461, at a 1/100 dilution, staining STAT1 (phospho S727) in paraffin embedded Human stomach adenocarcinoma tissue by Immunohistochemistry.
Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

SDS download

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Datasheet download

Download

发表研究结果有使用 ab109461？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab109461 被引用在 47 文献中.

Shen XY et al. Pre exposure to enriched environment alleviates brain injury after ischemia-reperfusion by inhibiting p38MAPK/STAT1 pathway. Mol Biol Rep 50:2243-2255 (2023). PubMed: 36572761
Wang X et al. Reduced Renal CSE/CBS/H2S Contributes to the Progress of Lupus Nephritis. Biology (Basel) 12:N/A (2023). PubMed: 36829595
Liu Z et al. IFN-α-2b Reduces Postoperative Arthrofibrosis in Rats by Inhibiting Fibroblast Proliferation and Migration through STAT1/p21 Signaling Pathway. Mediators Inflamm 2023:1699946 (2023). PubMed: 36915717
Lin R et al. Metformin attenuates diabetes-induced osteopenia in rats is associated with down-regulation of the RAGE-JAK2-STAT1 signal axis. J Orthop Translat 40:37-48 (2023). PubMed: 37304218
Zedan A et al. Antiviral Functions of Type I and Type III Interferons in the Olfactory Epithelium. Biomolecules 13:N/A (2023). PubMed: 38136633

View all Publications for this product

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Application

Western blot

Sample

Human Cell lysate - whole cell (HCT116)

Gel Running Conditions

Reduced Denaturing

Loading amount

10 µg

Specification

HCT116

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Chiara Vardabasso

Verified customer

提交于 Oct 27 2015

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

应用

The Abpromise guarantee

靶标

功能

疾病相关

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (47)

客户评价及客户问答

Western blot abreview for Anti-STAT1 (phospho S727) antibody [EPR3146]