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ERK1/2 (pT202/Y204 + Total) ELISA Kit (ab176660)

Key features and details

  • One-wash 90 minute protocol
  • Sample type: Cell Lysate, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

Overview

  • Product name

    ERK1/2 (pT202/Y204 + Total) ELISA Kit

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    (pT202/Y204) 6 3.3%
    (Total) 6 3%
    Inter-assay
    Sample n Mean SD CV%
    (pT202/Y204) 3 1.3%
    (Total) 3 4.9%

  • Sample type

    Cell Lysate, Tissue Homogenate

  • Assay type

    Semi-quantitative

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

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

  • Product overview

    Abcam’s ERK1/2 (pT202/Y204) and ERK1/2 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of ERK1/2 (pT202/Y204) and Total ERK1/2 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.


    ASSAY SPECIFICITY


    The ERK1/2 (pT202/Y204) assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737.2 [ERK1], NP 620407 [ERK2]) in cellular lysates, only when phosphorylated at Thr202/Tyr204.


    The ERK1/2 Total assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737.2 [ERK1], NP 620407 [ERK2]) in cellular lysates, irrespective of phosphorylation status.


    SPECIES REACTIVITY


    This kit detects ERK1/2 (pT202/Y204) and ERK1/2 Total in Human and mouse cell culture extracts. Detection in rat samples is also expected. Other species should be tested on a case-by-case basis.


    Serum and plasma samples have not been tested with this kit.


    As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.


     

  • Notes

    Estimated sensitivity: Phospho-ERK (Thr202/Tyr204): 0.1 ng/mL (tested with recombinant protein), Total ERK1/2: 0.6 ng/mL (tested with recombinant protein)
    Range: Phospho-ERK (Thr202/Tyr204): 0.2-20ng/mL, Total ERK1/2: 0.6-60 ng/mL

    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

Images

  • ERK1/2 (pT202/Y204 + Total) ELISA Kit (ab176660)
    ERK1/2 (pT202/Y204 + Total) ELISA Kit (ab176660)

    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 ERK1/2 (pT202/Y204) and ERK1/2 Total control lysate dilution series in 1X Cell Extraction Buffer PTR.
    Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 Total control lysate dilution series in 1X Cell Extraction Buffer PTR.

    The ERK1/2 lysate dilution series was prepared as described.  Raw data values are shown in the table.   

    Kit Control lysates are provided at a concentration that give consistent signal between different lots. Lysates are produced and formulated by signal intensity to be consistent to within 30% of the previous lot. As such, Control lysates are not provided with a protein concentration.

  • Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 Total control lysate dilution series in 1X Cell Extraction Buffer PTR.
    Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 Total control lysate dilution series in 1X Cell Extraction Buffer PTR.

    The ERK1/2 lysate dilution series was prepared as described.  Raw data values are shown in the table.   

    Kit Control lysates are provided at a concentration that give consistent signal between different lots. Lysates are produced and formulated by signal intensity to be consistent to within 30% of the previous lot. As such, Control lysates are not provided with a protein concentration.

  • Typical recombinant protein standard curve
    Typical recombinant protein standard curve

    Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 (Total) 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.

  • Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 Total recombinant protein standard curve.
    Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 Total recombinant protein standard curve.

    Raw data values are shown in the table.  Background-subtracted data values (mean +/- SD) are graphed.

  • Linearity of dilution
    Linearity of dilution

    Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of ERK1/2 (pT202/Y204) are normalized and plotted.

  • Comparison of ERK1/2 expression in different cell lines
    Comparison of ERK1/2 expression in different cell lines

    Cell line analysis for Total ERK1/2 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).

  • ERK1/2 (pT202/Y204) phosphorylation in response to EGF treatment
    ERK1/2 (pT202/Y204) phosphorylation in response to EGF treatment

    Induction of ERK1/2 (pT202/Y204) phosphorylation in MCF-7 cells in response to EGF treatment. MCF-7 cells were cultured in 96-well tissue culture plates, serum starved and treated (10 min) with a dose-range of EGF before cell lysis. Data from quadruplicate measurements of ERK1/2 (pT202/Y204) are plotted and compared against Total ERK1/2 protein levels. Comparative ERK1/2 (pT202/Y204) and ERK1/2 (Total) data also shown by Western Blot.

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