Overview

  • Product name

    JNK 1/2 (pT183/Y185) ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HEK extract 6 2.7%
    Inter-assay
    Sample n Mean SD CV%
    HEK extract 3 3.8%
  • Sample type

    Cell Lysate, Tissue Homogenate
  • Assay type

    Semi-quantitative
  • Sensitivity

    0.5 ng/ml
  • Range

    1 ng/ml - 100 ng/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 JNK1/2 (pT183/Y185) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of JNK1/2 (pT183/Y185) protein in Human and mouse 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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab176645 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.

     

  • Example of a typical JNK1/2 (pT183/Y185) cell lysate dilution series.  Background-subtracted data values (mean +/- SD) are graphed.

  • Example of a typical JNK1/2 (pT183/Y185) recombinant protein standard curve. The proportion of total protein that is phosphorylated is unknown - data is indicative only. Background-subtracted data values (mean +/- SD) are graphed.

  • Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1 x Cell Extraction Buffer PTR. Data from duplicate measurements of JNK1/2 (pT183/Y185) are normalized and plotted.

  • Cell line analysis for Total JNK1/2 from 200 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

  • Induction of JNK1/2 (pT183/Y185) phosphorylation in HeLa cells in response to anisomycin treatment. HeLa cells were cultured in 96-well tissue culture plates and treated (30 min) with a dose-range of anisomycin before cell lysis. Data from quadruplicate measurements of JNK1/2 (pT183/Y185) are plotted and compared against Total JNK1/2 protein levels. Comparative JNK1/2 (pT183/Y185) and JNK1/2 (Total) data also shown by Western Blot.

Protocols

References

This product has been referenced in:

  • Maayah ZH  et al. 2-Methoxyestradiol protects against pressure overload-induced left ventricular hypertrophy. Sci Rep 8:2780 (2018). Read more (PubMed: 29426916) »
See 1 Publication for this product

Customer reviews and Q&As

UNTESTED SPECIES/APP

Below average Average 3/5 (Ease of Use)
Abreviews
JNK1/2 ELISA Kit, ab176645
Sample- Rat tissue set #1
Absorbance-450nm
Microplate-96 wells
Microplate Reader –Spectramax Plus
Standard Curve
(Range 1.563ng/ml-100ng/ml)

Abcam user community

Verified customer

Submitted Jul 06 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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