Click-&-Go™ Plus 647 Labeling Kit
Click-&-Go™ Plus 647 Labeling Kit is a general purpose imaging kit that provides researchers everything required to perform the click reaction on alkyne tagged proteins with the far-red fluorescent AFDye 647 Picolyl Azide for subsequent downstream analysis. The kit utilizes a copper-chelating azide to raise the effective concentration of copper at the reaction site, which leads to dramatic acceleration of the rate of Cu(I)-catalyzed azide−alkyne cycloaddition (CuAAC) without the need to increase concentration of azide reagent and copper. Each kit includes sufficient material to perform least 30 cell or tissue assays based on a total reaction volume of 500 µL.
Click-&-Go™ Plus 647 Labeling Kit is a general purpose, high sensitivity imaging kit that provides researchers everything required to perform the click reaction on alkyne tagged proteins with the far-red fluorescent AFDye 647 Picolyl Azide for subsequent downstream analysis.
The kit utilizes an advanced fluorescent probe that incorporates a copper-chelating motif to raise the effective concentration of Cu(I) at the reaction site to dramatically improve the efficiency of the CuAAC reaction, resulting in a faster and more biocompatible CuAAC labeling (Figure 1). 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™ 647 Picolyl Azide is a water-soluble, pH-insensitive from pH 4 to pH 10, far-red-fluorescent probe with excitation ideally suited for the 633 nm or 647 nm laser lines. AFDye™ 647 is structurally similar to Alexa Fluor® 647, and spectrally is almost identical to Cy5 Dye, Alexa Fluor® 647, CF® 647 Dye, or any other Cyanine5 based fluorescent dyes.
Alexa Fluor® is a registered trademark of Thermo Fisher Scientific.
1. Jiang, H., et al. (2014). Monitoring Dynamic Glycosylation in Vivo Using Supersensitive Click Chemistry. Bioconjugate Chem.,, 25, 698-706. [PubMed]
2. 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]
3. 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 water-soluble ligand for copper-catalyzed azide-alkyne cycloadditions.
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AFDye 647 Picolyl Azide
Excitation maximum: 648 nm
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A newest generation, water-soluble accelerating ligand for CuAAC.
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