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Related Experiment Video

Updated: Jul 11, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP

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Published on: May 26, 2017

Development of EIA systems for active-form MAP kinase.

A Tani¹, M Noda, Y Ichimori

¹Discovery Research Laboratories III, Takeda Chemical Industries Ltd., 2-17-85 Juso-honmachi, Yodogawa-ku, Osaka, Japan. tani_akiyoshi@takeda.co.jp

Journal of Immunological Methods

|April 12, 2000

Summary

This summary is machine-generated.

New enzyme immunoassay (EIA) systems accurately measure active mitogen-activated protein kinases (MAPK), specifically ERK1 and ERK2. These assays offer enhanced specificity and convenience for studying cellular signaling pathways.

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Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Area of Science:

Biochemistry
Cell Biology
Immunology

Background:

Mitogen-activated protein kinases (MAPKs), including ERK1 and ERK2, are crucial signaling molecules.
Understanding the dynamic regulation of MAPK activity is essential for deciphering cellular responses to growth factors.

Purpose of the Study:

To develop novel quantitative sandwich enzyme immunoassay (EIA) systems for distinguishing active-form subtypes of p44 and p42 MAP kinase (ERK1 and ERK2).
To investigate the dynamic changes in ERK1 and ERK2 activity in response to platelet-derived growth factor (PDGF) and nerve growth factor (NGF).

Main Methods:

Development of EIA systems utilizing subtype-specific antibodies and a detector antibody targeting phosphorylated MAPKs.
Application of these EIA systems to analyze ERK1 and ERK2 activation in PDGF-treated rat mesangial cells and NGF-treated PC12 cells.

Main Results:

The developed EIA systems successfully distinguished between active-form ERK1 and ERK2.
Both ERK1 and ERK2 showed immediate activation upon stimulation, reaching maximum activity within 5-10 minutes.
Total MAPK activity measured by EIA correlated well with conventional methods.

Conclusions:

The novel EIA systems provide a highly specific, convenient, and efficient method for quantifying active ERK1 and ERK2.
These assays are applicable across multiple species (rat, human, mouse, rabbit), offering broad utility in various research investigations.
The systems facilitate the study of dynamic changes in MAPK signaling pathways.