Unit

UNIT 14.3 The Detection of MAPK Signaling

  1. Yoav Shaul,
  2. Rony Seger

Published Online: 1 OCT 2005

DOI: 10.1002/0471143030.cb1403s28

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Shaul, Y. and Seger, R. 2005. The Detection of MAPK Signaling. Current Protocols in Cell Biology. 28:14.3:14.3.1–14.3.34.

Author Information

  1. Department of Biological Regulation, The Weizmann Institute of Science, Rehovot, Israel

Publication History

  1. Published Online: 1 OCT 2005
  2. Published Print: SEP 2005

Abstract

Mitogen-activated protein kinase (MAPK) cascades are central pathways that participate in the intracellular transmission of extracellular signals. Each of the MAPK signaling cascades seems to consist of three to five tiers of protein kinases that sequentially activate each other by phosphorylation. Since the majority of MAPK cascade components are kinases, the methods used to detect their activation involve determining phosphorylation state and protein kinase activities. The Basic Protocol describes the use of immunoblotting with specific anti-phospho antibody to detect activation of MAPK components. Alternative methods described are immunoprecipitation of desired protein kinases followed by phosphorylation of specific substrates and the use of an in-gel kinase assay. These methods have proven useful in the study of the MAPK signaling cascades.

Keywords:

  • MAPK;
  • ERK;
  • JNK;
  • p38MAPK;
  • BMK;
  • protein kinases;
  • antibodies