title: Detection of ERK activation by a novel monoclonal antibody.
abstract: 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.
authors.affiliation: Department of Membrane Research and Biophysics, The Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Y
authors.initials: Y
authors.lastname: Yung
authors.forename: Y
authors.initials: Y
authors.lastname: Dolginov
authors.forename: Z
authors.initials: Z
authors.lastname: Yao
authors.forename: H
authors.initials: H
authors.lastname: Rubinfeld
authors.forename: D
authors.initials: D
authors.lastname: Michael
authors.forename: T
authors.initials: T
authors.lastname: Hanoch
authors.forename: E
authors.initials: E
authors.lastname: Roubini
authors.forename: Z
authors.initials: Z
authors.lastname: Lando
authors.forename: D
authors.initials: D
authors.lastname: Zharhary
authors.forename: R
authors.initials: R
authors.lastname: Seger
mesh_terms: D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D019556:COS Cells; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002460:Cell Line; D004331:Drosophila melanogaster; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology; D056890:Eukaryota; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010766:Phosphorylation; D017354:Point Mutation; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D015398:Signal Transduction; D018411:Spodoptera; D013912:Threonine; D014162:Transfection; D014443:Tyrosine
chemical_list: D000911:Antibodies, Monoclonal; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D013912:Threonine; D014443:Tyrosine; D004815:Epidermal Growth Factor; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020928:Mitogen-Activated Protein Kinases