Overview

  • Product nameSTAT3 (pY705) PhosphoTracer ELISA Kit
    See all STAT3 (phospho-Tyr705) kits
  • Detection methodFluorescent
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSemi-quantitative
  • Sensitivity
    50 µg protein/ml
  • Range
    50 µg protein/ml - 1000 µg protein/ml
  • Assay time
    2h 0m
  • Assay durationOne step assay
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Product overview

    PhosphoTracer assays use a traditional immuno-sandwich format, but with a major difference both the analyte and the assay reagents are added to the PhosphoTracer assay microplate at the same time. After a short incubation period, unbound assay reagents and analytes are washed away, and immuno-complexes containing both antibodies are detected. The process can take as little as 60 minutes to complete.

    PhosphoTracer kits also allow a higher degree of assay flexibility. In contrast to other ELISA formats, no antibodies are present on the assay microplate itself, so assays for several different targets can be performed in different wells on the same microplate. Simply mix the lysate with your target reagents of choice, using the microplate configuration of your choice.

    A whole new way of performing cellular assays, PhosphoTracer takes the hard work out of running a standard ELISA, while still giving the high quality results expected from a sandwich immunoassay. Fully self-contained kits are supplied in convenient 96-well packs. Simple to use and highly sensitive PhosphoTracer kits are designed to get results, fast.

     

    Abcam's PhosphoTracer STAT3 assay kits detect endogenous levels of the 2 splice variants, STAT3a and STAT3ß (Genbank Accessions NP_644805 and NP_998827) in cellular lysates. Phospho-STAT3 assay kits detect STAT3 only when phosphorylated at Tyr705. Total STAT3 assay kits detect STAT3 irrespective of phosphorylation status.



    The substrate used with the HRP conjugated detection antibody is a combination of 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP) (wavelength exc/em = 530-540nm / 590-600nm), a highly sensitive and stable substrate for HRP) and ADHP Dilution Buffer (a stabilized H2O2 solution). Learn more about the fluorogenic substrate, ADHP.

  • Tested applicationsSandwich ELISAmore details
  • PlatformMicroplate

Properties

  • FunctionTranscription 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 specificityHeart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • Involvement in diseaseDefects 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 similaritiesBelongs 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 localizationCytoplasm. 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
    • Stat3
    • STAT3_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab119650 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. Tested in HeLa, THP1, Jurkat, A431.

STAT3 (pY705) PhosphoTracer ELISA Kit images

  • As shown, using the STAT3 assay kits, a significant stimulation of STAT3 phosphorylation at Tyr705 is detected in A431 cells treated with EGF for 10 minutes compared with cells treated with AG1478, while a change in total STAT3 levels is also observed.
  • Day 1: A431 cells were seeded at 25K/well in a 96 well tissue culture plate in medium containing 10% FBS. Day 2: The cells were stimulated with various concentrations of EGF for 15 minutes. The media was removed, and the cells were lysed with 125 µL/well freshly prepared Lysis Mix, with shaking for 10 min. Lysates (50 µL) were transferred to replicate wells of a PhosphoTracer assay plate and analyzed for either phospho or total STAT3 using the standard PhosphoTracer protocol. Briefly, Antibody Mix specific for either phospho-STAT3, or Total STAT3 (50 µL/well), was added to the lysate, and the plate was incubated for 1 hr at room temp, with shaking. The plate was washed and Substrate Mix was added to the wells. The plate was covered in foil and incubated for 10 min with shaking. Signal in the wells was determined using a plate reader.

Protocols

References for STAT3 (pY705) PhosphoTracer ELISA Kit (ab119650)

ab119650 has not yet been referenced specifically in any publications.

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