| Development of EIA systems for active-form MAP kinase. | |
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MedLine Citation:
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PMID: 10758239 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Quantitative sandwich enzyme immunoassay (EIA) systems, that can distinguish between active-form subtypes of mitogen-activated protein kinases (p44 and p42 MAP kinase, also called ERK1 and ERK2), were developed employing subtype-specific antibodies as a solid phase and an antibody specific for the phosphorylated region of MAP kinases as the detector. Using these systems, we investigated the dynamic changes in the activity of ERK1 and ERK2 in platelet-derived growth factor (PDGF)-treated rat mesangial cells and nerve growth factor (NGF)-treated PC12. Both ERK1 and ERK2 were activated immediately after stimulation, and the activity reached a maximum at 5-10 min. The total activity of both subtypes correlated well with that obtained using the conventional method. Compared with the usual methods, these systems should have a higher specificity and be more convenient and suitable for experiments with multiple samples. Moreover, as these EIA systems can be applied not only to rat MAP kinases but also to human, mouse and rabbit MAP kinases, they are potentially very useful for a range of investigations. |
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Authors:
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A Tani; M Noda; Y Ichimori |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of immunological methods Volume: 238 ISSN: 0022-1759 ISO Abbreviation: J. Immunol. Methods Publication Date: 2000 Apr |
Date Detail:
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Created Date: 2000-06-06 Completed Date: 2000-06-06 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 1305440 Medline TA: J Immunol Methods Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 87-97 Citation Subset: IM |
Affiliation:
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Discovery Research Laboratories III, Takeda Chemical Industries Ltd., 2-17-85 Juso-honmachi, Yodogawa-ku, Osaka, Japan. tani_akiyoshi@takeda.co.jp |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Angiotensin II
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pharmacology Animals Antibody Specificity Benzimidazoles / pharmacology Cells, Cultured Enzyme Activation Glomerular Mesangium / cytology Humans Immunoenzyme Techniques / methods* Male Mice Mitogen-Activated Protein Kinase 1 / analysis*, immunology Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases / analysis*, immunology Muscle, Smooth, Vascular / cytology PC12 Cells Rabbits Rats Rats, Wistar Receptors, Angiotensin / antagonists & inhibitors Tetrazoles / pharmacology |
| Chemical | |
Reg. No./Substance:
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0/Benzimidazoles; 0/Receptors, Angiotensin; 0/Tetrazoles; 11128-99-7/Angiotensin II; 139481-59-7/candesartan; EC 2.7.11.24/Mitogen-Activated Protein Kinase 1; EC 2.7.11.24/Mitogen-Activated Protein Kinase 3; EC 2.7.11.24/Mitogen-Activated Protein Kinases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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