title: Labeling a protein with fluorophores using NHS ester derivitization.
abstract: N-hydroxysuccinimde (NHS) ester-mediated derivitization involves the reaction of this amine-reactive group with the primary amines of a protein or a biomolecule. Using NHS chemistry allows one to conjugate various fluorescent probes, biotin, and cross-linkers to primary amines. For example, we use NHS ester chemistry to fluorescently label the amino terminus of a protein with the dye, 5-(and-6)-carboxyfluorescein, succinimidyl ester (5(6)-FAM, SE).
authors.affiliation: Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Jagpreet S
authors.initials: JS
authors.lastname: Nanda
authors.affiliation: Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: jlorsch@jhmi.edu.
authors.forename: Jon R
authors.initials: JR
authors.lastname: Lorsch
mesh_terms: D002021:Buffers; D004952:Esters; D005452:Fluoresceins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012996:Solutions; D013056:Spectrophotometry, Ultraviolet; D013194:Staining and Labeling; D013388:Succinimides / Q000737:chemistry*
chemical_list: D002021:Buffers; D004952:Esters; D005452:Fluoresceins; D005456:Fluorescent Dyes; D011506:Proteins; D012996:Solutions; D013388:Succinimides; C024098:6-carboxyfluorescein
keywords: Efficiency of labeling calculation; Fluorescent labeling of protein; Fluorophores; Lyophilized protein; NHS ester derivitization; UV spectrophotometer