UNIT 18.4 Analysis of Phosphorylation of Unlabeled Proteins
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- 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.