title: A capture coupling method for the covalent immobilization of hexahistidine tagged proteins for surface plasmon resonance.
abstract: Surface plasmon resonance (SPR) is a robust method to detect and quantify macromolecular interactions; however, to measure binding interactions, one component must be immobilized on a sensor surface. This is typically achieved using covalent immobilization via free amines or thiols, or noncovalent immobilization using high-affinity interactions such as biotin/streptavidin or antibody/antigen. In this chapter we describe a robust method to covalently immobilize His(6) fusion proteins on the sensor surface for SPR analysis.
authors.affiliation: Department of Pharmacology, The University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Kimple
authors.forename: Robin E
authors.initials: RE
authors.lastname: Muller
authors.forename: David P
authors.initials: DP
authors.lastname: Siderovski
authors.forename: Francis S
authors.initials: FS
authors.lastname: Willard
mesh_terms: D000588:Amines / Q000737:chemistry; D002021:Buffers; D019204:GTP-Binding Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006639:Histidine / Q000378:metabolism*; D055767:Immobilized Proteins / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D000588:Amines; D002021:Buffers; C471213:His-His-His-His-His-His; D055767:Immobilized Proteins; D009842:Oligopeptides; D006639:Histidine; D019204:GTP-Binding Proteins