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    stat1-py701--total-stat1-elisa-kit-ab126457.pdf

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Microbiology Interspecies Interaction Host Virus Interaction
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STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)

  • Datasheet
  • SDS
  • Protocol Booklet
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Western blot - STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
  • Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
  • Western blot - STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
  • Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
  • Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)

Key features and details

  • Sample type: Cell Lysate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

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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-pan STAT1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and STAT1 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-STAT1 (Tyr701) antibody is added to detect phosphorylated STAT1. In the remaining wells, biotinylated anti-STAT1 antibody is used to detect pan STAT1. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin is pipetted into 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 pan STAT1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • 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 10ml
    5X Assay Diluent 1 x 15ml
    HRP-conjugated anti-rabbit IgG 500X concentrate 1 x 25µl
    HRP-Streptavidin Concentrate 1 x 200µl
    Pan Detection Antibody STAT1: biotinylated anti-pan STAT1 1 vial
    Pan STAT1 Microplate (12 strips x 8 wells) coated with anti-pan STAT1 antibody 1 unit
    Phospho Detection Antibody STAT1 (Tyr701) 1 vial
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • STAT family
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Signal transduction proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Phosphoprotein ELISA kits
  • 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.
  • 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
  • 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
    • Transcription factor ISGF3 components p91/p84
    • XStat1
    see all
  • Database links

    • Entrez Gene: 6772 Human
    • Entrez Gene: 20846 Mouse
    • Omim: 600555 Human
    • SwissProt: P42224 Human
    • SwissProt: P42225 Mouse
    • Unigene: 642990 Human
    • Unigene: 743244 Human
    • Unigene: 277406 Mouse

    Associated products

      Images

      • Western blot - STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
        Western blot - STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
        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).
      • Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
        Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
        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.
      • Western blot - STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
        Western blot - STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
        A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using Western Blot.
      • Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
        Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
        A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
      • Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
        Sandwich ELISA - STAT1 (pY701) + total STAT1 Human ELISA Kit (ab126457)
        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

      • Protocol Booklet

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
      • SDS
    • References (2)

      Publishing research using ab126457? Please let us know so that we can cite the reference in this datasheet.

      ab126457 has been referenced in 2 publications.

      • Torbica T  et al. Chronic Inflammation in Response to Injury: Retention of Myeloid Cells in Injured Tissue Is Driven by Myeloid Cell Intrinsic Factors. J Invest Dermatol 139:1583-1592 (2019). PubMed: 30703358
      • 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). PubMed: 26709698

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-2 of 2 Abreviews or Q&A

      Question

      The homogenate/lysis is cloudy, will this affect the reaction? Using RAW264.7 cells might sonication be required?

      Read More

      Abcam community

      Verified customer

      Asked on Apr 09 2013

      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.

      Read More

      Abcam Scientific Support

      Answered on Apr 09 2013

      Question

      Can thid kit be used with tissue lysates? If so is there any protocol variations that are required?

      Read More

      Abcam community

      Verified customer

      Asked on Oct 15 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Oct 15 2012

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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