AAT Bioquest

Click Chemistry Building Blocks

Reaction scheme of Alkyne with a dye azide
The reaction scheme of a biomolecule alkyne with a dye azide.
"Click Chemistry" is a term introduced by K. B. Sharpless in 2001 to describe reactions that are high in yields, wide in scope, and create only by-products that can be removed without chromatography. Click chemistry reactions are stereospecific, simple to perform and can be conducted in easily removable or benign solvents. This concept was developed in parallel with the interest within the pharmaceutical, material, and other industries in capabilities of generating large libraries of compounds for screening in discovery research. Several types of reaction have been identified that fulfill these criteria. They are thermodynamically-favored reactions that lead specifically to one product, such as nucleophilic ring opening reactions of epoxides and aziridines; non-aldol type carbonyl reactions, such as formation of hydrazones and heterocycles; eletrophilic additions to carbon-carbon multiple bonds, such as oxidative formation of epoxides and Michael Additions; and cycloaddition reactions.

The reaction scheme of a biomolecule azide with a dye alkyne.
The reaction scheme of a biomolecule azide with a dye alkyne.
An examination of the azide-alkyne cycloaddition shows that it fulfills many of the prerequisites. The copper-catalyzed azide"alkyne cyloaddition is a two-step process. First, one reaction partner-either an azide or alkyne linked to a "building block" such as a peptide or an oligonucleotide, is incorporated by conventional synthesis. Subsequently, the other reaction partner-the complementary alkyne or azide linked to a fluorescent dye, biotin or other detection reagent-is "clicked" into place in the presence of catalytic copper (I). One reaction partner must be an azide derivative and the other an alkyne derivative, but functional moiety can serve as either the incorporated molecule or the detection molecule. The reaction is also regiospecific, yielding exclusively 1,4-disubstituted-1,2,3-triazole linkages. The 1,2,3-triazole linkage between a peptide or an oligonucleotide and a dye is extremely stable. It is not susceptible to hydrolysis, oxidation or reduction. AAT Bioquest offers a variety of dye azides and alkynes for labeling peptides and oligonucleotides. These clickable reagents include both common fluorescent dyes (e.g., fluoresceins, rhodamines and cyanines) and non-fluorescent quenchers. Our Tide Fluor™ and Tide Quencher™ dyes are specifically optimized for preparing novel FRET substrates.

Table 1. Dye Azides for Labeling Oligonucleotides and Peptides

Cat No.
Product Name
Ex (nm)
Em (nm)
EC (cm-1M-1)
Unit Size
508AMCA Azide35345518,0000.183 0.1531 mg
143Cyanine 3 azide [equivalent to Cy3® azide]555565150,0000.0420.0731 mg
153Cyanine 5 azide [equivalent to Cy5® azide]649665250,0000.0260.0301 mg
178Cyanine 5.5 azide [equivalent to Cy5.5® azide]678701230,0000.0940.1011 mg
163Cyanine 7 azide [equivalent to Cy7® azide]749776275,0000.0250.0361 mg
1315-FAM Azide49452175,0000.2470.17810 mg
1336-FAM Azide49451875,0000.2550.17210 mg
2406-HEX azide53355074,0000.3000.1275 mg
2486-JOE azide52054873,0000.3260.2175 mg
4946-ROX azide57560295,0000.3070.1795 mg
4865-TAMRA azide54757375,0000.2880.1785 mg
4906-TAMRA azide54757375,0000.3350.1805 mg
2446-TET azide52153676,0000.1910.1045 mg
484Texas Red® azide *Single Isomer*58860195,0000.4560.3605 mg
2236Tide Fluor™ 1 azide [TF1 azide]34544220,0000.1830.1875 mg
2252Tide Fluor™ 2 azide [TF2 azide]50052775,0000.2880.0911 mg
2254Tide Fluor™ 3 azide [TF3 azide]55558475,0000.3310.1781 mg
2300Tide Fluor™ 4 azide [TF4 azide]59061390,0000.4890.4361 mg
2275Tide Fluor™ 5WS azide [TF5WS azide]649664250,0000.0230.0271 mg
2302Tide Fluor™ 6WS azide [TF6WS azide]676695220,0000.1110.1011 mg
2304Tide Fluor™ 7WS azide [TF7WS azide]749775275,0000.0090.0491 mg
2188Tide Quencher™ 1 azide [TQ1 azide]490 20,0000.1470.1945 mg
2211Tide Quencher™ 2 azide [TQ2 azide]501 21,0000.1000.1205 mg
2231Tide Quencher™ 3 azide [TQ3 azide]573 22,0000.0850.0915 mg
2068Tide Quencher™ 4WS azide [TQ4WS azide]603 90,0000.1490.136 1 mg
2082Tide Quencher™ 5WS azide [TQ5WS azide]661 130,0000.0720.0821 mg
2097Tide Quencher™ 6WS azide [TQ6WS azide]704 130,0000.1200.1021 mg
2112Tide Quencher™ 7WS azide [TQ7WS azide]763 140,0000.0720.0911 mg

Table 2. Dye Alkynes for Labeling Oligonucleotides and Peptides

Cat No.
Product Name
Ex (nm)
Em (nm)
Unit Size
507AMCA Alkyne3534551 mg
144Cyanine 3 alkyne [equivalent to Cy3® alkyne]5555651 mg
154Cyanine 5 alkyne [equivalent to Cy5® alkyne]6496651 mg
179Cyanine 5.5 alkyne [equivalent to Cy5.5® alkyne]6787011 mg
164Cyanine 7 alkyne [equivalent to Cy7® alkyne]7497761 mg
1325-FAM Alkyne49451810 mg
1346-FAM Alkyne49451810 mg
2416-HEX alkyne5335505 mg
2496-JOE alkyne5205485 mg
4956-ROX alkyne5756025 mg
4875-TAMRA alkyne5475735 mg
4916-TAMRA alkyne5475735 mg
2456-TET alkyne5215365 mg
485Texas Red® alkyne *Single Isomer*5886015 mg
485Texas Red® alkyne *Single Isomer*5886015 mg
2237Tide Fluor™ 1 alkyne [TF1 alkyne]3454425 mg
2253Tide Fluor™ 2 alkyne [TF2 alkyne]5005271 mg
2255Tide Fluor™ 3 alkyne [TF3 alkyne]5555841 mg
2301Tide Fluor™ 4 alkyne [TF4 alkyne]5906181 mg
2276Tide Fluor™ 5WS alkyne [TF5WS alkyne]6496641 mg
2303Tide Fluor™ 6WS alkyne [TF6WS alkyne]6766951 mg
2305Tide Fluor™ 7WS alkyne [TF7WS alkyne]7497751 mg
2189Tide Quencher™ 1 alkyne [TQ1 alkyne]490 5 mg
2212Tide Quencher™ 2 alkyne [TQ2 alkyne]501 5 mg
2232Tide Quencher™ 3 alkyne [TQ3 alkyne]573 5 mg
2069Tide Quencher™ 3 alkyne [TQ3 alkyne]603 1 mg
2069Tide Quencher™ 4WS alkyne [TQ4WS alkyne]603 1 mg
2083Tide Quencher™ 5WS alkyne [TQ5WS alkyne]661 1 mg
2098Tide Quencher™ 6WS alkyne [TQ6WS alkyne]704 1 mg
2113Tide Quencher™ 7WS alkyne [TQ7WS alkyne]763 1 mg