Introduction
Click Chemistry is a new approach to rapidly and selectively react ("click") a pair of functional groups with each other in mild, aqueous conditions. Click Chemistry reactions can be categorized into two separate groups: (1) Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC); (2) Strain-promoted Azide-Alkyne Click Chemistry reaction (SPAAC).
Biomolecules that have been functionalized with a CLICK moiety (e.g. a terminal Alkyne or Azide) can be detected by labeling with a compatible CLICKable fluorescent dye (e.g. a fluorescent Azide or Alkyne) that allows the subsequent visualization by fluorescence spectroscopy. ABP Biosciences has developed a series of CLICKable biotin and fluorescent dyes for efficient “click” labeling of biomolecules.
Features
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Highly selective, low background labeling
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Rapid and quantitative labeling
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Multiple fluorophore selection for multicolor imaging
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Andy Fluor™ dye for brighter signal
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Figure 1. Click chemistry labeling
Standard CuAAC reactions
Azides and alkynes can be used interchangeably in standard CuAAC reactions, so either one can serve as the tagged substrate or be labeled for the detection step. Some biomolecules are sensitive to the amount of copper used to catalyze standard click reactions. In particular, proteins such as GFP and R-PE exhibit reduced fluorescence and nucleic acids can be denatured by a high-copper environment.
DBCO alkynes for copper-free reactions
DBCO reagents can be used to perform click reactions with azide-modified targets without the use of heavy metal catalysis. DBCO reactions are ideal for surface labeling of live cells and also minimize damage to fluorescent proteins like GFP or R-PE.
CLICKable Biotin and Fluorescent Dye Selection Guide