Dye affinity chromatography

Current Protocols in Protein Science
E Stellwagen

Abstract

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.

References

Jun 1, 1990·Journal of Molecular Recognition : JMR·C R LoweS Eapen
Jan 1, 1990·Methods in Enzymology·E Stellwagen
Oct 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·J B RobinsonE Stellwagen
Apr 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·J B RobinsonE Stellwagen
Oct 5, 1987·Biotechnology and Bioengineering·Y D ClonisC R Lowe

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