Overview

  • Product name

    c-Jun pS73 ELISA Kit
    See all c-Jun kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HeLa 6 2.9%
    Inter-assay
    Sample n Mean SD CV%
    HeLa 3 8.2%
  • Sample type

    Cell culture extracts, Cell Lysate
  • Assay type

    Semi-quantitative
  • Sensitivity

    10 µg/ml
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    Abcam’s c-Jun (pS73) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (ab205706) is designed for the semi-quantitative measurement of c-Jun (pS73) protein in Human and mouse cells.


    c-Jun is a component of the transcription factor AP-1, which is composed of dimers of members of the Fos and Jun protein families. c-Jun transcriptional activity is regulated by phosphorylation at Ser63 and Ser73. Many factors, including mitogenic and stress-induced signaling pathways, stimulate phosphorylation of c-Jun at Ser63 and Ser73 and activate c-Jun/AP-1-dependent transcription. JNK, whose activity is stimulated by the same signals, binds to the amino-terminal region of c-Jun and phosphorylates it directly. c-Jun/AP-1 dependant transcription is involved in the regulation of genes involved in many diverse and sometimes opposing cellular processes, including proliferation, survival, apoptosis, metastasis and inflammation, depending on the cell type.


     


    An alternative fluorescent substrate, ADHP, can be used with this assay. A microplate reader capable of measuring fluorescence is required if using this product.

  • Notes

    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

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab205706 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

  • 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.

     

  • Cell line analysis for c-Jun 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).

  • Activation of c-Jun (pS73) phosphorylation and total c-Jun expression in HeLa cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates and treated (30 minutes) with a dose-range of TNFα before cell lysis. Data from triplicate measurements of c-Jun (pS73) are plotted and compared against total c-Jun protein levels.

  • Example of a typical c-Jun (pS73) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

Protocols

References

ab205706 has not yet been referenced specifically in any publications.

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