Overview

  • Product name
    STAT3 (pY705) + total STAT3 ELISA Kit
    See all STAT3 (phospho Tyr705) + total STAT3 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, Rat, Human
  • Product overview

    STAT3 (pY705) + total STAT3 ELISA Kit (ab126459) 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 STAT3 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 phospho-STAT3 (Tyr705) and total STAT3 in human, mouse and rat cell lysates (help normalize the results of phospho-STAT3 from different cell lysate being compared). A pan STAT3 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and STAT3 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phospho-STAT3 (Tyr705) is used to detect phosphorylated STAT3 or biotinylated anti-total-STAT3 is used to detect total STAT3. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-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 STAT3 (Tyr705) or total STAT3 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Get results in 90 minutes with STAT3 (pY705 + Total) ELISA Kit (ab176666) from our SimpleStep ELISA® range.

  • 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 lysate buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    80x HRP-Streptavidin Concentrate 1 x 200µl
    Detection Antibody STAT3 1 vial
    Detection Antibody STAT3 (Tyr705) 1 vial
    HRP-conjugated anti-rabbit IgG 1 x 25µl
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    STAT3 Microplate (12 strips x 8 wells) coated with anti-pan STAT3 antibody 1 unit
    Stop Solution 1 x 8ml
    TMB Substrate Reagent 1 x 12ml
  • 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
    • HIES
    • Signal transducer and activator of transcription 3
    • signal transducer and activator of transcription 3 (acute-phase response factor)
    • Stat3
    • STAT3_HUMAN
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab126459 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

  • 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.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 20 min. Cell lysates were analyzed using this phosphoELISA.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 20 min. Cell lysates were analyzed using Western Blot.
  • The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes. Serial dilutions of lysates were analyzed in this ELISA.
  • 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-STAT3 (Tyr705).

Protocols

References

This product has been referenced in:

See 1 Publication for this product

Customer reviews and Q&As

Thank you for contacting us.

The tissue samples are yet to be tested with this kit so we cannot share the data how well this kit will perform. As this kit is ELISA so we don't see any reason why this should not be working with tissues lysates....

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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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