Overview

  • Product name
    STAT3 (Tyr705) In-Cell ELISA Kit
  • Detection method
    Colorimetric
  • Sample type
    Adherent cells
  • Assay type
    Cell-based (qualitative)
  • Assay time
    5h 10m
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    ab126427 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of STAT3 (Tyr705) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human, mouse and rat cell lines. By determining STAT3 protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot.

    In the STAT3 (Tyr705) In-Cell ELISA Kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, Anti-Phospho-STAT3 (Tyr705) or Anti-STAT3 is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-mouse IgG is added to the wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. 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: In-Cell ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    Anti-Mouse IgG Concentrate (Item I) 1 x 10µl
    5X Blocking Buffer Concentrate (Item F) 1 x 20ml
    Fixing Solution (Item D) 1 x 30ml
    Uncoated 96-well Microplate (Item A) 1 unit
    Mouse Anti-Phospho-STAT3 (Tyr705) Concentrate (Item G) 1 x 10µl
    Mouse Anti-STAT3 Concentrate (Item H) 1 x 10µl
    Quenching Buffer Concentrate (30x) (Item E) 1 x 2ml
    Stop Solution 1 x 14ml
    TMB Substrate Reagent 1 x 12ml
    20X Wash Buffer A Concentrate (Item B) 1 x 30ml
    20X Wash Buffer B Concentrate (Item C) 1 x 30ml
  • Research areas
  • Function
    Transcription factor that binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA.
  • Tissue specificity
    Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • Involvement in disease
    Defects in STAT3 are the cause of hyperimmunoglobulin E recurrent infection syndrome autosomal dominant (AD-HIES) [MIM:147060]; also known as hyper-IgE syndrome or Job syndrome. AD-HIES is a rare disorder of immunity and connective tissue characterized by immunodeficiency, chronic eczema, recurrent Staphylococcal infections, increased serum IgE, eosinophilia, distinctive coarse facial appearance, abnormal dentition, hyperextensibility of the joints, and bone fractures.
  • Sequence similarities
    Belongs to the transcription factor STAT family.
    Contains 1 SH2 domain.
  • Post-translational
    modifications
    Tyrosine phosphorylated upon stimulation with EGF (By similarity). Tyrosine phosphorylated in response to IL-6, IL-11, CNTF, LIF, CSF-1, EGF, PDGF, IFN-alpha and OSM. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus.
  • Cellular localization
    Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3.
  • Information by UniProt
  • Alternative names
    • Acute-phase response factor
    • APRF
    • DNA binding protein APRF
    • Signal transducer and activator of transcription 3
    • STAT 3
    • Stat3
    • STAT3_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab126427 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
In-Cell ELISA Use at an assay dependent concentration.

Images

  • A431 cells were stimulated by different concentrations of EGF for 10 minutes at 37°C.

  • A431 cells were stimulated by different concentrations of EGF for 30 minutes at 37°C.

  • Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-Stat3 (Tyr705) and Anti-Stat3 antibodies were used in both detection assays.
  • A431 cells were stimulated by different concentrations of EGF for 10 minutes at 37°C.
  • A431 cells were stimulated by different concentrations of EGF for 30 minutes at 37°C.

Protocols

References

ab126427 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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