STAT1 (pY701) ELISA Kit (ab126456)
- Product nameSTAT1 (pY701) ELISA KitSee all STAT1 (phospho Tyr701) kits ...
- Detection methodColorimetric
- Tests1 x 96 well plate
- Sample typeCell Lysate
- Assay typeSandwich (qualitative)
- Assay time5h 00m
- Assay durationMultiple steps standard assay
- Species reactivityReacts with: Mouse, Human
- Product overview
ab126456 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human and mouse cell lysates. By determining phosphorylated STAT1 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of Human and Mouse phospho-STAT1 (Tyr701). An anti-phospho-STAT1 (Tyr701) antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated STAT1 (Tyr701) present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated STAT1 antibody is used to detect phosphorylated STAT1. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of STAT1 (Tyr701) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
- Tested applicationsSandwich ELISA more details
- Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 x 96 tests 200X HRP-Streptavidin Concentrate 1 x 200µl 20X Wash Buffer Concentrate 1 x 25ml 2X Cell Lysate Buffer 1 x 5ml 5X Assay Diluent 1 x 15ml Detection Antibody STAT1: biotinylated anti-STAT1 2 vials Positive Control: lyophilized powder from A431 cell lysate 1 vial STAT1 Microplate (12 strips x 8 wells) coated with anti-phosphoSTAT1 (Tyr701) antibody 1 unit Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml
- 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.
- 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.
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.
- Cellular localizationCytoplasm. Nucleus. Translocated into the nucleus in response to IFN-gamma-induced tyrosine phosphorylation and dimerization.
- ISGF 3
- Signal transducer and activator of transcription 1
- Signal transducer and activator of transcription 1 91kDa
- Signal transducer and activator of transcription 1-alpha/beta
- Transcription factor ISGF-3 components p91/p84
Our Abpromise guarantee covers the use of ab126456 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
STAT1 (pY701) ELISA Kit images
A431 cells were treated with recombinant Human EGF at 37°C for 20 min. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed using ab126456.
A431 cells were treated or untreated with 100 ng/ml recombinant Human EGF for 10 min. Cell lysates were analyzed using ab126456.
A431 cells were treated or untreated with 100 ng/ml recombinant Human EGF for 10 min. Cell lysates were analyzed by Western Blot.
A431 cells were treated with 100 ng/mL recombinant Human EGF for 20 minutes to induce phosphorylation of EGF R. Serial dilutions of lysates were analyzed using ab126456.
The A431 cells were treated with 100 ng/mL recombinant Human EGF for 20 minutes to induce phosphorylation of EGF R. Serial dilutions of lysates were analyzed by western blot. Immunoblots were incubated with anti-phospho-STAT1 (Tyr701).
References for STAT1 (pY701) ELISA Kit (ab126456)
ab126456 has not yet been referenced specifically in any publications.