Introduction
This note describes how to label proteins of MW > 10000 using
MSD® TAG-NHS-Ester. The procedure involves a 1 hour incubation
step followed by a simple column separation to isolate the
labeled protein.
MSD TAG-NHS-Ester, or Ruthenium (II) tris-bipyridine, Nhydroxysuccinimide
is an amine-reactive label often used for
detection in assays formatted with MSD technology (Figure 1). Nhydroxysuccinimide
esters readily couple to primary amine
groups of proteins to form a stable amide bond. The reaction is
favored at slightly alkaline pH in which a nucleophilic attack by
an unprotonated amine on an ester results in amide bond
formation. The reaction is rapid and occurs under mild
conditions.
MSD TAG-labeled conjugates are stable and may be used at
low concentrations. These features minimize time, costs, and
labor since large, stable batches of conjugate can be prepared,
validated, and used for long periods of time. MSD TAG-NHSEster
is a popular conjugate for labeling molecules that contain
primary amines (e.g., lysines in proteins) since the method is
fast and easy.
关键字： [Ru(dppz)(dmb′-CO2-);
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