Click chemistry describes a class of chemical reactions that use bio-orthogonal or biologically unique moieties to label and detect a molecule of interest in mild, aqueous conditions. The click reaction involves a copper-catalyzed triazole formation from an azide and an alkyne. The azide and alkyne moieties can be used interchangeably; either one can be used to tag the molecule of interest, while the other is used for subsequent detection.

The biotin alkyne is reactive with azide via a copper-catalyzed click reaction. Biotin can be subsequently detected with streptavidin, avidin or NeutrAvidin® biotin-binding protein.

Efficiency—the click reaction is complete in less than 1 hour;

Specificity—the reaction between the label and detection tag is selective and specific;

Stability—the reaction product contains an irreversible, covalent bond;

Biologically inert—the components of the reaction do not undergo any side reactions.

1.Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins.

Clark PM, Dweck JF, Mason DE, Hart CR, Buck SB, Peters EC, Agnew BJ, Hsieh-Wilson LC,J Am Chem Soc (2008) 130:11576-11577

2.Robust fluorescent detection of protein fatty-acylation with chemical reporters.

Charron G, Zhang MM, Yount JS, Wilson J, Raghavan AS, Shamir E, Hang HC,J Am Chem Soc (2009) 131:4967-4975

3.Regulation of calcium/calmodulin-dependent kinase IV by O-GlcNAc modification.

Dias WB, Cheung WD, Wang Z, Hart GW,J Biol Chem (2009) 284:21327-21337