Human STAT5 (Tyr694) In-Cell ELISA Kit (ab126429)

Overview

  • Product name
    Human STAT5 (Tyr694) 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: Human
  • Product overview

    ab126429 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 STAT5 (Tyr694) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human cell lines. By determining STAT5 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 In-Cell STAT5 (Tyr694) 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-STAT5 (Tyr694) or Anti-STAT5 (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-mouse IgG (secondary antibody) 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

Applications

Our Abpromise guarantee covers the use of ab126429 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-STAT5 (Tyr694) and STAT5 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

ab126429 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

We have not tried the assay with non-adherent cells and we are not aware of any other lab trying this either. It might be accomplished by centrifuging the cells in a 96-well plate after each wash, to pull them down into pellets in each well, but you wo...

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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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