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Labeling a protein with fluorophores using NHS ester derivitization.

Methods in enzymology (2014-01-16)
Jagpreet S Nanda, Jon R Lorsch
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).

MATERIALS
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Product Description

Sigma-Aldrich
5(6)-Carboxyfluorescein, suitable for fluorescence, BioReagent, ≥95% (HPLC)