UNIT 18.4 Analysis of Phosphorylation of Unlabeled Proteins

  1. Bartholomew M. Sefton

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1804s40

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Sefton, B. M. 2001. Analysis of Phosphorylation of Unlabeled Proteins. Current Protocols in Molecular Biology. 40:18.4:18.4.1–18.4.7.

Author Information

  1. The Salk Institute, San Diego, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1997


Phosphorylation of unlabeled proteins can be detected using immunological or enzymatic techniques. Anti-phosphotyrosine antibodies are used with immunoblots to detect tyrosine phosphorylation. This unit provides a protocol for detection of antibodies by 125I-labeled protein A or enhanced chemiluminescence (ECL). Detection of enzymatic dephosphorylation using a general or phosphoamino acid-specific phosphatase and subsequent SDS-PAGE mobility shift (or a change in activity) is also described.