UNIT 9.2 Dye Affinity Chromatography

  1. Earle Stellwagen

Published Online: 1 MAY 2001

DOI: 10.1002/0471140864.ps0902s00

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Stellwagen, E. 2001. Dye Affinity Chromatography. Current Protocols in Protein Science. 00:9.2:9.2.1–9.2.16.

Author Information

  1. University of Iowa, Iowa City, Iowa

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1995


Dye affinity chromatography is a protein purification procedure based on the high affinity of immobilized dyes for the binding sites on many proteins. It is a rapid, inexpensive, and versatile method that is applicable to the purification of crude cellular extracts. This unit presents protocol for the three types of dye affinity chromatography: negative chromatography, positive chromatography, and tandem chromatography. An initial protocol describes a chromatographic procedure in which a small volume of the protein mixture to be purified is applied to a series of miniature columns, each containing a different immobilized dye. Analysis of the flowthrough and bound material allows determination of the optimum dye material for larger-scale purification. An alternate procedure describes a similar initial selection procedure using centrifugation instead of chromatography. A support protocol describes a simple procedure for immobilization of free dyes.