SimpleStep

ERK1/2 (pT202/Y204) ELISA Kit (ab176640)

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Other - ERK1/2 (pT202/Y204) ELISA Kit (ab176640)
  • Typical cell lysate dilution series
  • Typical recombinant protein standard curve
  • Linearity of dilution
  • Comparison of total ERK1/2 expression in different cell lines
  • ERK1/2 (pT202/Y204) phosphorylation in response to EGF treatment.

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 0.1 ng/ml
  • Range: 0.2 ng/ml - 20 ng/ml
  • Sample type: Cell Lysate, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

Overview

  • Product name

    ERK1/2 (pT202/Y204) ELISA Kit
    See all ERK1 + ERK2 kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    A431 extract 6 3.3%
    Inter-assay
    Sample n Mean SD CV%
    A431 extract 3 1.3%

  • Sample type

    Cell Lysate, Tissue Homogenate

  • Assay type

    Semi-quantitative

  • Sensitivity

    0.1 ng/ml

  • Range

    0.2 ng/ml - 20 ng/ml

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

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

  • Product overview

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


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

  • Notes

    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 - ERK1/2 (pT202/Y204) ELISA Kit (ab176640)
    Other - ERK1/2 (pT202/Y204) ELISA Kit (ab176640)

    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 ERK1/2 (pT202/Y204) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

  • Typical recombinant protein standard curve
    Typical recombinant protein standard curve

    Example of a typical ERK1/2 (pT202/Y204) 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
    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 total ERK1/2 expression in different cell lines
    Comparison of total 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.

References (6)

Publishing research using ab176640? Please let us know so that we can cite the reference in this datasheet.

ab176640 has been referenced in 6 publications.

  • Xu Y  et al. miR-133b affects cell proliferation, invasion and chemosensitivity in renal cell carcinoma by inhibiting the ERK signaling pathway. Mol Med Rep 22:67-76 (2020). PubMed: 32377748
  • Maayah ZH  et al. 2-Methoxyestradiol protects against pressure overload-induced left ventricular hypertrophy. Sci Rep 8:2780 (2018). PubMed: 29426916
  • Vallejo A  et al. NF?B Inhibition Mitigates Serum Amyloid A-Induced Pro-Atherogenic Responses in Endothelial Cells and Leukocyte Adhesion and Adverse Changes to Endothelium Function in Isolated Aorta. Int J Mol Sci 20:N/A (2018). PubMed: 30597899
  • Schaefer TL  et al. Acamprosate in a mouse model of fragile X syndrome: modulation of spontaneous cortical activity, ERK1/2 activation, locomotor behavior, and anxiety. J Neurodev Disord 9:6 (2017). ELISA ; Mouse . PubMed: 28616095
  • Bukong TN  et al. Therapeutic Benefits of Spleen Tyrosine Kinase Inhibitor Administration on Binge Drinking-Induced Alcoholic Liver Injury, Steatosis, and Inflammation in Mice. Alcohol Clin Exp Res N/A:N/A (2016). Sandwich ELISA ; Mouse . PubMed: 27177528
  • Sager HB  et al. Proliferation and Recruitment Contribute to Myocardial Macrophage Expansion in Chronic Heart Failure. Circ Res 119:853-64 (2016). Sandwich ELISA ; Mouse . PubMed: 27444755

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p-ERK stimulation in HEK cell lysates

 Good Good 4/5 (Ease of Use)
Abreviews
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This kit was used to assess the induction of ERK phosphorylation in vitro in HEK cells lysates.
Cells were treated with PMA (0.050 ng/ml) in the presence or abscence of PD98059 (50 uM). Cells were briefly washed with ice cold PBS and lysated. After centrifugation, the supernatant was used as recommended by the kit booklet. The kit was easy to use and would recommend it.

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Submitted Sep 10 2018

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