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Azide-containing Trifunctional Reagents

TAMRA-Azide-Biotin is a trifunctional biotinylation reagent that is suitable for the simultaneous labeling of terminal Alkyne- or strained Alkyne (e.g. DBCO = ADIBO)-tagged biomolecules with both a Biotin and a Fluorescent moiety (Fig. 1).

The Azide moiety (1st function) enables the attachment to terminal Alkyne- or strained Alkyne (e.g. DBCO)-tagged molecules via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively. The Biotin moiety (2nd function) allows the subsequent affinity purification/immobilization of the labeled molecule with Streptavidin Agarose. Optimal streptavidin binding is ensured by the introduction of hydrophilic PEG spacers that increase the water solubility and reduce steric hindrance. The carboxytetramethylrhodamine (TAMRA) fluorophore (3rd function) can be used for the sensitive detection of the immobilized protein.

Figure 1: Chemical structure of TAMRA-Azide-Biotin. 1. Azide moiety, 2. Biotin moiety, 3. TAMRA moiety.