Unit

UNIT 14.2 Immunological Detection of Phosphorylation

  1. Zhong Yao,
  2. Rony Seger

Published Online: 1 MAY 2001

DOI: 10.1002/0471143030.cb1402s00

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Yao, Z. and Seger, R. 2001. Immunological Detection of Phosphorylation. Current Protocols in Cell Biology. 00:14.2:14.2.1–14.2.15.

Author Information

  1. The Weizmann Institute of Science, Rehovot, Israel

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1999

Abstract

Incorporation of phosphates into serine, threonine, and tyrosine acceptors in proteins is a common mechanism for regulating protein function. This unit presents protocols that use specific anti-phosphoamino acid (PAA) and anti-phosphoprotein antibodies to detect protein phosphorylation and protein kinase activity. Immunoblotting to detect protein phosphorylation using either anti-PAA or anti-phosphoprotein antibodies. This is a convenient method that usually yields impressive results. Phosphorylation can also be detected by immunoprecipitation followed by immunoblot analysis or by immunofluorescent staining; these methods are typically more complicated and time consuming. All three methods have been successfully used to detect protein phosphorylation with a wide variety of antibodies and most phosphorylated proteins.