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    stat1-ps727--total-stat1-elisa-kit-ab126455.pdf

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Microbiology Interspecies Interaction Host Virus Interaction
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STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (1)References (1)

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Western blot - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
  • Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
  • Western blot - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
  • Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
  • Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)

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 (pS727) + 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

    ab126455 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 phospho-STAT1 (Ser727) and total STAT1 (help normalize the results of phospho-STAT1 from different cell lysate being compared). An anti-STAT1 (Ser727) (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 (Ser727) 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.

  • 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 5ml
    5X Assay Diluent 1 x 15ml
    Detection Antibody anti-pan STAT1 2 vials
    Detection Antibody anti-phospho STAT1 (Ser727) 1 vial
    500x HRP-conjugated anti-rabbit IgG concentrate 1 x 25ml
    HRP-Streptavidin Concentrate 2 x 200µl
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    STAT1 Microplate with anti-pan STAT1 antibody 1 unit
    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 (pS727) + total STAT1 ELISA Kit (ab126455)
        Western blot - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes. Serial dilutions of lysates were analyzed by Western blot. Immunoblots were incubated with anti-phospho-STAT1 (Ser727).
      • Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes. Serial dilutions of lysates were analyzed in this ELISA.
      • Western blot - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        Western blot - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed by Western Blot.
      • Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        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 (pS727) + total STAT1 ELISA Kit (ab126455)
        Sandwich ELISA - STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
        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 (1)

      Publishing research using ab126455? Please let us know so that we can cite the reference in this datasheet.

      ab126455 has been referenced in 1 publication.

      • 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

      Question

      I have used ab126455 to detect total and phospho serine STAT1; interestingly the OD values using the positive control lysate and my own samples show higer values detected for phospho STAT1 than for total STAT1;
      Here are some examples of OD values derived from the positive control samples:
      OD values
      phospho total
      P1 2.9 2.6
      P2 2.3 2.0
      P3 1.1 0.7

      Read More

      Abcam community

      Verified customer

      Asked on Sep 12 2013

      Answer

      Thank you for your inquiry. This is entirely possible because two different capture antibodies are being used with different binding affinities, sensitivities, and specificities. Keep in mind that this assay is semi-quantitative (i.e. not measuring actual levels or concentrations) so you are only measuring relative protein expression between like targets (i.e. you can only compare “like vs. like”).


      So a higher OD from a phospho result does not necessarily mean that there is more phosphorylated protein than total protein in that sample (this of course would not make sense). All this would really tell you is that the anti-phospho antibody had a higher affinity for the protein than the anti-pan had for the protein. In brief, you can only compare “total vs. total” and “phospho vs. phospho”
      So in general this kit is useful to compare total and phospho STAT1 between samples, not to determine the percentage of total STAT1 protein that is phosphorylated.

      Read More

      Jeremy Kasanov

      Abcam Scientific Support

      Answered on Sep 12 2013

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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