Overview

  • Product name

    STAT1 (pY701) + total STAT1 ELISA Kit
    See all STAT1 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell Lysate
  • Assay type

    Semi-quantitative
  • Assay time

    5h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse, Human
  • Product overview

    ab126457 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in 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 Blot analysis.

    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of Human and Mouse phospho-STAT1 (Tyr701) and total STAT1 (help normalize the results of phospho-STAT1 from different cell lysate being compared). An anti-STAT1 (Tyr701) (half plate, red marker on left side) and anti-total STAT1 antibody (half plate, black marker on right side) has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated (left side) and total (right side) STAT1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated STAT1 is used to detect phosphorylated or total 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) or total STAT1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell Lysis Buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    Detection Antibody STAT1: biotinylated anti-STAT1 2 vials
    HRP-Streptavidin Concentrate 1 x 200µl
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    STAT1 Microplate (12 strips x 8 wells) coated with anti-phospho-STAT1 (Y701) + anti-STAT1 antibody 1 unit
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • Function

    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.
  • Involvement in disease

    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.
  • Sequence similarities

    Belongs to the transcription factor STAT family.
    Contains 1 SH2 domain.
  • Post-translational
    modifications

    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.
  • Cellular localization

    Cytoplasm. Nucleus. Translocated into the nucleus in response to IFN-gamma-induced tyrosine phosphorylation and dimerization.
  • Information by UniProt
  • Alternative names

    • Signal transducer and activator of transcription 1 91kD
    • CANDF7
    • DKFZp686B04100
    • IMD31A
    • IMD31B
    • IMD31C
    • ISGF 3
    • ISGF-3
    • OTTHUMP00000163552
    • OTTHUMP00000165046
    • OTTHUMP00000165047
    • OTTHUMP00000205845
    • Signal transducer and activator of transcription 1
    • Signal transducer and activator of transcription 1 91kDa
    • Signal transducer and activator of transcription 1, 91kD
    • Signal transducer and activator of transcription 1-alpha/beta
    • STAT 1
    • Stat1
    • STAT1_HUMAN
    • STAT91
    • Transcription factor ISGF 3 components p91 p84
    • Transcription factor ISGF-3 components p91/p84
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab126457 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

  • The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of STAT1. Serial dilutions of lysates were analyzed by Western blot. Immunoblots were incubated with anti-phospho-STAT1 (Tyr701).
  • The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of STAT1. Serial dilutions of lysates were analyzed in this ELISA.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using Western Blot.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
  • 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 in this ELISA.

Protocols

References

This product has been referenced in:

  • Kroetz DN  et al. Type I Interferon Induced Epigenetic Regulation of Macrophages Suppresses Innate and Adaptive Immunity in Acute Respiratory Viral Infection. PLoS Pathog 11:e1005338 (2015). Read more (PubMed: 26709698) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

We recommend centrifuging the cloudy homogenates and collect the supernatant for testing purposes. Sonication is a fine method for homogenization. Make sure you gently mixes the collected sample by pipetting up and down before testing the ELISA.

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Answer

Thank you for contacting Abcam.

I have talked to the lab and this kit can be used for cell and tissue lysates. You can use the buffer provided to lyse the samples or follow the guidelines in the attachment. The volume required will depend on the amount of tissue you are using.

If there is anything else I can help you with, please let me know.

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