FastClick™ Digoxigenin (DIG) Azide
FastClick™ Digoxigenin (DIG) Azide contains both the CAG moiety of FastClick (for assisting click efficiency) and DIG hapten (as the detection tag) for developing DIG-based probes. It readily reacts with an alkyne-containing biomolecule under extremely mild conditions. DIG is a steroid found exclusively in the flowers and leaves of the plants Digitalis purpurea, Digitalis orientalis and Digitalis lanata. It is a commonly used hapten (a small molecule with high antigenicity) that finds utilities in biological detections similarly to other popular haptens such as 2,4-Dinitrophenol (DNP) and biotin. DIG conjugates and tags are widely used in fluorescence imaging, fluorescence in situ hybridization (FISH) and other nucleic acid detections. FastClick™ reagents have been developed by the scientists of AAT Bioquest for enhancing the yield and reaction speed of copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. They contain a copper-chelating ligand that significantly stabilizes the Cu(I) oxidation state and thus accelerates the click reaction. They do not require the use of an external copper-chelator (such as the common THPTA or BTTAA). The high concentration of copper chelators is known to have a detrimental effect on DNA/RNA, thus causing biocompatibility issues. The introduction of a copper-chelating moiety at the reporter molecule allows for a dramatic raise of the effective Cu(I) concentration at the reaction site and thus accelerates the reaction. Under extremely mild conditions the FastClick™ azides and alkynes react much faster in high yield compared to the corresponding conventional CuAAC reactions. Click chemistry was developed by K. Barry Sharpless as a robust and specific method of ligating two molecules together. Two important characteristics make click chemistry attractive for assembling biomolecules. First, click reactions are bio-orthogonal, thus the click chemistry-functionalized biomolecules would not react with the natural biomolecules that lack a clickable functional group. Second, the reactions proceed with ease under mild conditions, such as at room temperature and in aqueous media.
![The reaction (Green Bar) of FastClick Cy5 Azide with coumarin alkyne occurs under extremely mild conditions (e.g., [Azide] = 0.02 mM, [Alkyne] = 0.02 mM, [CuSO4] = 0.02 mM, [Sodium Ascorbate] = 5 mM, in 100 mM HEPES) under which the common Cy5 azide does not effectively react with the coumarin alkyne substrate.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Ffastclick-digoxigenin-dig-azide%2Ffigure-for-fastclick-digoxigenin-dig-azide_anY0M.png&w=128&q=25)
References
View all 2 references: Citation Explorer
Analysis of a temperature-sensitive mutation in Uba1: Effects of the click reaction on subsequent immunolabeling of proteins involved in DNA replication.
Authors: Sugaya, Kimihiko and Ishihara, Yoshie and Inoue, Sonoe
Journal: FEBS open bio (2015): 167-74
Authors: Sugaya, Kimihiko and Ishihara, Yoshie and Inoue, Sonoe
Journal: FEBS open bio (2015): 167-74
Cell surface display yields evolvable, clickable antibody fragments.
Authors: Van Deventer, James A and Yuet, Kai P and Yoo, Tae Hyeon and Tirrell, David A
Journal: Chembiochem : a European journal of chemical biology (2014): 1777-81
Authors: Van Deventer, James A and Yuet, Kai P and Yoo, Tae Hyeon and Tirrell, David A
Journal: Chembiochem : a European journal of chemical biology (2014): 1777-81