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Detection of ERK activation by a novel monoclonal antibody
Author(s) -
Yung Yuval,
Dolginov Yakov,
Zhong Yao,
Rubinfeld Hadara,
Michael Dan,
Hanoch Tamar,
Roubini Eli,
Lando Zeev,
Zharhary Dorit,
Seger Rony
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00442-0
Subject(s) - mapk/erk pathway , monoclonal antibody , phosphorylation , kinase , microbiology and biotechnology , chemistry , antibody , western blot , protein kinase a , biochemistry , biology , immunology , gene
The mitogen‐activated protein kinase, ERK is activated by a dual phosphorylation on threonine and tyrosine residues. Using a synthetic diphospho peptide, we have generated a monoclonal antibody directed to the active ERK. The antibody specifically identified the active doubly phosphorylated, but not the inactive mono‐ or non‐ phosphorylated forms of ERKs. A direct correlation was observed between ERK activity and the intensity in Western blot of mitogen‐activated protein kinases from several species. The antibody was proven suitable for immunofluorescence staining, revealing a transient reactivity with ERKs that were translocated to the nucleus upon stimulation. In conclusion, the antibody can serve as a useful tool in the study of ERK signaling in a wide variety of organisms.