AFDye 680 Picolyl Azide
AFDye™ 680 Picolyl Azide is an advanced fluorescent probe that incorporates a copper-chelating motif to raise the effective concentration of Cu(I) at the reaction site to boost the efficiency of the CuAAC reaction, resulting in a faster and more biocompatible CuAAC labeling. Up to 40-fold increase of signal intensity, compared to conventional azides, was reported (see Selected References).
AFDye™ 680 Picolyl Azide is an advanced fluorescent probe that incorporates a copper-chelating motif to raise the effective concentration of Cu(I) at the reaction site to boost the efficiency of the CuAAC reaction, resulting in a faster and more biocompatible CuAAC labeling. Up to 40-fold increase of signal intensity, compared to conventional azides, was reported (see Selected References).
In addition, the use picolyl azides instead of conventional azides allows for at least a tenfold reduction in the concentration of the copper catalyst without sacrificing the efficiency of labeling, significantly improving biocompatibility of CuAAC labeling protocol.
In summary, the introduction of a copper-chelating motif into azide probe leads to a substantial increase in the sensitivity and reduced cell toxicity of CuAAC detection alkyne-tagged biomolecules. This will be of special value for the detection of low abundance targets or living system imaging.
AFDye 680 (Alexa Fluor ® 680 equivalent) is a bright and photostable near-IR dye that is spectrally identical to Alexa Fluor® 680, IRDye® 680RD and Cy5.5. AFDye 680 is often used for small animal imaging applications in the 700 nm wavelength and provides excellent clearance profiles in animals. Flow cytometry is another common application for this dye. AFDye 680 ideally suited for the 633 nm laser line. This near-IR fluorescent dye is water soluble and pH-insensitive from pH 4 to pH 10. The long wavelength emission allows for detection in complex samples with auto-fluorescent background signals.
- Jiang, H., et al. (2014). Monitoring Dynamic Glycosylation in Vivo Using Supersensitive Click Chemistry. Bioconjugate Chem.,, 25, 698-706. [PubMed]
- Uttamapinant, C., et al. (2012). Fast, Cell-Compatible Click Chemistry with Copper-Chelating Azides for Biomolecular Labeling. Angew. Chem. Int. Ed,., 51, 5852-56. [PubMed]
- Gaebler, A.,et al. (2016). A highly sensitive protocol for microscopy of alkyne lipids and fluorescently tagged or immunostained proteins. J. Lipid. Res., 57, 1934-47. [PubMed]
A general purpose click chemistry reaction buffer kit.
An alkyne analog of puromycin that is efficiently incorporated into newly translated proteins in complete methionine-containing media.
Alkyne-containing, noncanonical amino acid.