Document Detail


Development of EIA systems for active-form MAP kinase.
MedLine Citation:
PMID:  10758239     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
A Tani; M Noda; Y Ichimori
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of immunological methods     Volume:  238     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  2000 Apr 
Date Detail:
Created Date:  2000-06-06     Completed Date:  2000-06-06     Revised Date:  2013-05-28    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  87-97     Citation Subset:  IM    
Affiliation:
Discovery Research Laboratories III, Takeda Chemical Industries Ltd., 2-17-85 Juso-honmachi, Yodogawa-ku, Osaka, Japan. tani_akiyoshi@takeda.co.jp
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MeSH Terms
Descriptor/Qualifier:
Angiotensin II / pharmacology
Angiotensin Receptor Antagonists
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
Tetrazoles / pharmacology
Chemical
Reg. No./Substance:
0/Angiotensin Receptor Antagonists; 0/Benzimidazoles; 0/Tetrazoles; 11128-99-7/Angiotensin II; 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; S8Q36MD2XX/candesartan

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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