Unit

UNIT 13.6 Preparation and Application of Polyclonal and Monoclonal Sequence-Specific Anti-Phosphoamino Acid Antibodies

  1. Tong Sun,
  2. Ralph B. Arlinghaus

Published Online: 1 FEB 2004

DOI: 10.1002/0471140864.ps1306s34

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Sun, T. and Arlinghaus, R. B. 2004. Preparation and Application of Polyclonal and Monoclonal Sequence-Specific Anti-Phosphoamino Acid Antibodies. Current Protocols in Protein Science. 34:13.6:13.6.1–13.6.27.

Author Information

  1. University of Texas M.D. Anderson Cancer Center, Houston, Texas

Publication History

  1. Published Online: 1 FEB 2004
  2. Published Print: NOV 2003

Abstract

This unit discusses the issues that must be considered in the design, production, and characterization of polyclonal and monoclonal sequence-specific anti-phosphoamino acid antibodies. Protocols are provided for generating and purifying such antibodies, and methods are also provided for producing useful polyclonal antibodies in a non-purified form. Support protocols describe coupling of peptides or phosphotyrosine to a solid support for use in affinity chromatography. An example of the generation, purification, and characterization of two sequence-specific anti-phosphopeptide antibodies specific for different sequences of a single phosphoprotein is described. The cross-reactivity of such antibodies, which is a common problem with anti-peptide antibodies, is also discussed.