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Use of Tandem Affinity Chromatography for Purification of Cannabinoid Receptor CB2

  • Silvia C. Locatelli-Hoops
  • Alexei A. YeliseevEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1177)

Abstract

Tandem affinity purification has been increasingly applied to isolation of recombinant proteins. It relies on two consecutive chromatographic steps that take advantage of the affinity tags placed at opposing ends of the target protein. This allows for efficient removal of contaminating proteins, including products of proteolytic degradation of the fusion that lack either N- or C-terminal tags. Here, we describe the use of two small affinity tags, a poly-histidine tag and a Strep-tag for expression and purification of the human cannabinoid receptor CB2, an integral membrane G protein-coupled receptor.

Key words

His-tag Strep-tag Dual affinity tags Membrane protein CB2 receptor StrepTactin Ni-NTA 

Notes

Acknowledgments

This work was supported by the Intramural Research Program of the National Institute on Alcoholism and Alcohol Abuse, National Institutes of Health.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Silvia C. Locatelli-Hoops
    • 1
  • Alexei A. Yeliseev
    • 1
    Email author
  1. 1.Laboratory of Membrane Biochemistry and BiophysicsNational Institute on Alcohol Abuse and Alcoholism, National Institutes of HealthBethesdaUSA

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