title: Use of tandem affinity chromatography for purification of cannabinoid receptor CB₂.
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.
authors.affiliation: Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, 5625 Fishers Lane, Bethesda, MD, 20892, USA.
authors.forename: Silvia C
authors.initials: SC
authors.lastname: Locatelli-Hoops
authors.forename: Alexei A
authors.initials: AA
authors.lastname: Yeliseev
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli; D006639:Histidine / Q000737:chemistry; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D009842:Oligopeptides / Q000737:chemistry; D043885:Receptor, Cannabinoid, CB2 / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: C099517:Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly; D009842:Oligopeptides; D043885:Receptor, Cannabinoid, CB2; C033223:polyhistidine; D006639:Histidine