2. Academic Validation
  3. Labeling a protein with fluorophores using NHS ester derivitization

Labeling a protein with fluorophores using NHS ester derivitization

  • Methods Enzymol. 2014;536:87-94. doi: 10.1016/B978-0-12-420070-8.00008-8.
Jagpreet S Nanda 1,  Jon R Lorsch 2
Affiliations

Affiliations

  • 1 Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
  • 2 Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: jlorsch@jhmi.edu.
PMID: 24423269 DOI: 10.1016/B978-0-12-420070-8.00008-8
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).

Keywords

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

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