STAT1 (pY701) ELISA Kit (ab176653)
- Datasheet
- References
- Protocols
Overview
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Product nameSTAT1 (pY701) ELISA Kit
See all STAT1 kits -
Detection methodColorimetric
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Precision
Intra-assay Sample n Mean SD CV% HeLa extract 6 5.8% Inter-assay Sample n Mean SD CV% HeLa extract 3 5.2% -
Sample typeCell Lysate, Tissue Homogenate
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Assay typeSemi-quantitative
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Sensitivity< 20 µg/ml
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Range20 µg/ml - 2000 µg/ml
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Assay time1h 30m
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Assay durationOne step assay
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Species reactivityReacts with: Human
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Product overview
Abcam’s STAT1 (pY701) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of STAT1 (pY701) protein in Human cells.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
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Tested applicationsSuitable for: Sandwich ELISAmore details
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PlatformMicroplate
Properties
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Storage instructionsStore at +4°C. Please refer to protocols.
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Components 1 x 96 tests 10X Wash Buffer PT 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 12ml Lyophilized STAT1 Control Lysate 1 vial Plate Seal 1 unit SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit STAT1 (pY701) Capture Antibody 1 x 3ml STAT1 (pY701) Detector Antibody 1 x 3ml Stop Solution 1 x 12ml TMB Substrate 1 x 12ml -
Research areas
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FunctionSignal 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.
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Involvement in diseaseNote=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. -
Sequence similaritiesBelongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
Post-translational
modificationsPhosphorylated 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. -
Cellular localizationCytoplasm. Nucleus. Translocated into the nucleus in response to IFN-gamma-induced tyrosine phosphorylation and dimerization.
- Information by UniProt
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Alternative names
- Signal transducer and activator of transcription 1 91kD
- CANDF7
- DKFZp686B04100
see all -
Database links
- Entrez Gene: 6772 Human
- Omim: 600555 Human
- SwissProt: P42224 Human
- Unigene: 642990 Human
- Unigene: 743244 Human
Associated products
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SimpleStep ELISA kits
Applications
Our Abpromise guarantee covers the use of ab176653 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Typical cell lysate dilution seriesExample of a typical STAT1 (pY701) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Linearity of dilutionLinearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1 x Cell Extraction Buffer PTR. Data from duplicate measurements of STAT1 (pY701) are normalized and plotted.
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STAT1 (pY701) phosphorylation in response to IFN? treatmentInduction of STAT1 (pY701) phosphorylation in HeLa cells in response to IFN-gamma treatment. HeLa cells were cultured in 96-well tissue culture plates, serum-starved and treated (20 min) with a dose-range of IFN-gamma before cell lysis. Data from quadruplicate measurements of STAT1 (pY701) are plotted. Comparative STAT1 (pY701) and STAT1 (Total) data also shown by Western Blot.
Protocols
References
ab176653 has not yet been referenced specifically in any publications.
