Abstract
Publisher Summary This chapter discusses the purification of calmodulin by Ca 2+ -dependent affinity chromatography. The chapter describes the use of phenothiazine conjugates in calmodulin purification. There are four proven purification steps that can be used effectively for preparing extracts for affinity chromatography: heat treatment, ammonium sulfate fractionation, diethylaminoethyl (DEAE) batch step, and trichloroacetic acid precipitation. Calmodulin have been purified from many different sources representing vertebrates, invertebrates, plants, and fungi. There is no unique combination of these steps that can function universally for the preparation of extracts for affinity chromatography. Each tissue or organism tends to have unique purification problems and for these reasons, this chapter presents an outline of the protocols that are usually applicable for use with phenothiazine affinity chromatography. Frequently, many calmodulin samples eluted from phenothiazine conjugates have other contaminants. Therefore, it is necessary to use a final purification step to remove these contaminants. Ion-exchange steps usually work well for achieving final purification.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.