Detection of ERK activation by a novel monoclonal antibody

FEBS Letters

Roubini, Eli¹  Yung, Yuval²  Dolginov, Yakov¹  Zhong, Yao²  Lando, Zeev¹  Michael, Dan³  Rubinfeld, Hadara²  Seger, Rony²  Hanoch, Tamar²  Zharhary, Dorit¹ 

[1] Sigma Israel Chemicals Ltd., Rehovot, Israel;Department of Membrane Research and Biophysics, The Weizmann Institute of Science, Rehovot 76100, Israel;Department of Pharmacology, New York University, New York, USA

关键词: MAPK;    Signal transduction;    Monoclonal antibody;    Cellular localization;   

DOI  :  10.1016/S0014-5793(97)00442-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown