Introduction to click chemistry: a new method for the labeling and modification of biomolecules
Click chemistry is a reaction between azide and alkyne yielding covalent product – 1,5-disubstituted 1,2,3-triazole. This process is also known as a cooper-catalyzed alkyne-azide cycloaddition or CuAAC.
Click chemistry is based on copper catalysis, and the catalyst is often introduced as a cooper-ligand complex.
Among the wide variety of organic reactions, click chemistry has been selected as a conjugation chemistry reaction because of several advantages.
- It is very selective. The click chemistry reaction takes place only between azide and alkyne components. It does not interfere with most other organic groups present in DNA and proteins being labeled, such as amino and carboxy groups.
- There are no azides and alkynes in native biomolecules. These groups should be specially introduced into DNA and proteins. Alkyne-containing DNA can be prepared with alkyne phosphoramidite during standard oligo synthesis. Proteins labeled with azide and alkyne can be made using azide-activated ester and alkyne-activated ester.
- Click chemistry takes place in water. Aqueous DMSO, DMF, acetonitrile, alcohols, or pure water and buffers can be used for the reaction. The reaction is biocompatible and can take place in living cells.
- The reaction is quick and quantitative. Click chemistry is a tool that allows the preparation of nanomoles of conjugates in diluted solutions.
- The reaction is pH-insensitive. Unlike the reaction of NHS-esters with amines and some other conjugation chemistries, the reaction mixture does not need to control pH. There is no need to add any specific buffer, acid, or base – click chemistry works well in pH intervals of 4-11.
- The protocol is simple! For an example, see our recommended DNA labeling protocol.
Click chemistry thus became a tool for the universal modification of DNA, proteins, conjugate preparation, and fluorescent labeling. This is where Lumiprobe comes to help – we provide reagents and protocols for the facile and efficient synthesis of diverse azido- and alkyne-labeled biomolecules, as well as reactive fluorescent dyes and other reporter groups. With these reagents, you can easily prepare conjugates in your Lab.
Here are just several examples:
Fluorescently labeled oligonucleotides & dual-labeled probes for real-time PCR
We provide alkyne phosphoramidites to easily synthesize alkyne-modified oligos and then label them with fluorescent dye azides. Based on click chemistry, this technology provides significant advantages over labeling via activated esters or fluorescent dye amidites.
Fluorescent & biotinylated DNA
Use alkynyl triphosphates to incorporate alkyne in DNA by PCR, termination, or nick-translation. You can, after that, label DNA with any dye or biotin in your lab without the need for specifically labeled triphosphates!
Fluorescent peptides, proteins, and antibodies
We provide alkyne and azide-activated esters for modifying proteins and peptides with either azide or alkyne. You can use alkyne- or azido-modified proteins to prepare conjugates with DNA, reporter molecules, and solid surfaces.
Peptide-oligonucleotide conjugates
We provide azido activated ester for labeling peptides and alkyne amidites for synthesizing ethynylated oligos.
Biomolecules immobilized on nearly any solid phase
We would be glad to consult you on modifying solid surfaces, providing custom solid phases, and guiding you to success!
Click chemistry auxiliary reagents & catalysts are available in our catalogue...
... and nearly any other conjugates you can imagine can be done by click chemistry... Contact us to find how click chemistry can help you!
Some reviews on DNA modification with click chemistry:
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