Detection of ERK activation by a novel monoclonal antibody

FEBS Lett. 1997 May 26;408(3):292-6. doi: 10.1016/s0014-5793(97)00442-0.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Antibodies, Monoclonal*
  • Antibody Specificity
  • Binding Sites, Antibody
  • Blotting, Western
  • COS Cells
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Cell Line
  • Drosophila melanogaster
  • Enzyme Activation
  • Epidermal Growth Factor / pharmacology
  • Eukaryota
  • HeLa Cells
  • Humans
  • Kinetics
  • Mice
  • Mitogen-Activated Protein Kinases / metabolism*
  • Mutagenesis, Site-Directed
  • Nerve Tissue Proteins / metabolism*
  • Phosphorylation
  • Point Mutation
  • Rats
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Signal Transduction
  • Spodoptera
  • Threonine
  • Transfection
  • Tyrosine


  • Antibodies, Monoclonal
  • Nerve Tissue Proteins
  • Recombinant Proteins
  • Threonine
  • Tyrosine
  • Epidermal Growth Factor
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Mitogen-Activated Protein Kinases