Labeling a protein with fluorophores using NHS ester derivitization

Jagpreet S. Nanda, Jon R. Lorsch

Research output: Chapter in Book/Report/Conference proceedingChapter

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).

Original languageEnglish (US)
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationProtein Part A
PublisherAcademic Press Inc.
Pages87-94
Number of pages8
ISBN (Print)9780124200708
DOIs
StatePublished - Jan 1 2014

Publication series

NameMethods in Enzymology
Volume536
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Efficiency of labeling calculation
  • Fluorescent labeling of protein
  • Fluorophores
  • Lyophilized protein
  • NHS ester derivitization
  • UV spectrophotometer

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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