SimpleStep

STAT5 A/B pY694/699 ELISA Kit (ab176656)

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Other - STAT5 A/B pY694/699 ELISA Kit (ab176656)
  • Typical cell lysate dilution series
  • Induction of STAT5 A/B (pY694/699) phosphorylation in A431 cells in response to EGF treatment.
  • Cell line analysis for Total STAT5 A/B

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 5 µg/ml
  • Range: 10 µg/ml - 1000 µg/ml
  • Sample type: Adherent cells, Cell culture extracts, Suspension cells, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Human

Overview

  • Product name

    STAT5 A/B pY694/699 ELISA Kit
    See all STAT5 kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    A431 extract 6 2.8%
    Inter-assay
    Sample n Mean SD CV%
    A431 extract 3 4%

  • Sample type

    Cell culture extracts, Adherent cells, Suspension cells, Tissue Homogenate

  • Assay type

    Semi-quantitative

  • Sensitivity

    < 5 µg/ml

  • Range

    10 µg/ml - 1000 µg/ml

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Product overview

    Abcam’s STAT5 A/B (pY694/699) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of STAT5 A/B (pY694/699) protein in Human cells.


    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm

  • Notes

    Abcam’s STAT5 A/B (pY694/699) SimpleStep ELISA kit (ab176656) has been re-developed with new detector antibody.  From February 18, 2016 the kit will be delivered with a new detector total STAT5 antibody. This change was necessary because of a decrease in the performance of the polyclonal STAT5 antibody that was previously used for detection in recent production lots. We have identified a new antibody to use in the SimpleStep ELISA platform that provides higher performance.

    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

Associated products

    Images

    • Other - STAT5 A/B pY694/699 ELISA Kit (ab176656)
      Other - STAT5 A/B pY694/699 ELISA Kit (ab176656)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Typical cell lysate dilution series
      Typical cell lysate dilution series

      Example of a typical STAT5 A/B (pY694/699) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

    • Induction of STAT5 A/B (pY694/699) phosphorylation in A431 cells in response to EGF treatment.
      Induction of STAT5 A/B (pY694/699) phosphorylation in A431 cells in response to EGF treatment.

      A431 cells were cultured in 96-well tissue culture plates and treated (15 min) with a dose-range of EGF before cell lysis. Data from quadruplicate measurements of STAT5 A/B (pY694/699) are plotted and compared against Total STAT5 A/B levels.

    • Cell line analysis for Total STAT5 A/B
      Cell line analysis for Total STAT5 A/B

      Cell line analysis for Total STAT5 A/B from 100 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

    References (0)

    Publishing research using ab176656? Please let us know so that we can cite the reference in this datasheet.

    ab176656 has not yet been referenced specifically in any publications.

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