AAT Bioquest

FastClick™ 5-FAM Azide

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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.
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Physical properties
Molecular weight588.58
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
Alternative formats
FastClick™ 5-FAM Alkyne
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FastClick™ 6-FAM Alkyne
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Show More (43)


See also: Click Chemistry
Molecular weight
FastClick™ 5-FAM Azide contains both the CAG moiety of FastClick (for assisting click efficiency) and 5-FAM fluorophore (as the fluorescence tag) for developing 5-FAM-based fluorescent probes. 5-FAM is one of the most widely used green fluorophores, in particular, for labeling peptides. It has almost the identical fluorescence spectra to Alexa Fluor 488, a rhodamine 110 analog. 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.


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of FastClick™ 5-FAM Azide to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM169.9 µL849.502 µL1.699 mL8.495 mL16.99 mL
5 mM33.98 µL169.9 µL339.801 µL1.699 mL3.398 mL
10 mM16.99 µL84.95 µL169.9 µL849.502 µL1.699 mL

Molarity calculator

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View all 9 references: Citation Explorer
Programmed immobilization of living cells using independent click pairs.
Authors: Zhu, Chengyuan and Takemoto, Hiroyasu and Higuchi, Yuriko and Yamashita, Fumiyoshi
Journal: Biochemical and biophysical research communications (2024): 149556
Concerning the photophysics of fluorophores towards tailored bioimaging compounds: a case study involving S100A9 inflammation markers.
Authors: Steiner, Simon T and Maisuls, Iván and Junker, Anna and Fritz, Günter and Faust, Andreas and Strassert, Cristian A
Journal: Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society (2023): 2093-2104
The development of potent, competitive CXCR4 antagonists for the prevention of cancer metastasis.
Authors: Hamshaw, Isabel and Cominetti, Marco M D and Lai, Wing-Yee and Searcey, Mark and Mueller, Anja
Journal: Biochemical pharmacology (2023): 115921
Manipulating Diastereomeric Bicyclononynes to Sensitively Determine Enzyme Activity and Facilitate Macromolecule Conjugations.
Authors: Huang, Chien-Hui and Hou, Sin-Yu and Severance, Scott and Hwang, Chi-Ching and Fang, Bo-Kai and Gong, Ming-Mao and Yu, Siao-Lei and Weng, Yung-Chieh and Wang, Li-Fang and Dai, Chia-Yen and Wang, Su-Hung and Kuo, Hsing-Tao and Wang, Jeh-Jeng and Wang, Tzu-Pin
Journal: ACS omega (2023): 46073-46090
Design and Synthesis of Fluorophore-Tagged Disparlure Enantiomers to Study Pheromone Enantiomer Discrimination in the Pheromone-Binding Proteins from the Gypsy Moth, Lymantria dispar.
Authors: Pinnelli, Govardhana R and Plettner, Erika
Journal: Journal of chemical ecology (2021)
Click Chemistry on the Surface of Ultrasmall Gold Nanoparticles (2 nm) for Covalent Ligand Attachment Followed by NMR Spectroscopy.
Authors: van der Meer, Selina Beatrice and Loza, Kateryna and Wey, Karolin and Heggen, Marc and Beuck, Christine and Bayer, Peter and Epple, Matthias
Journal: Langmuir : the ACS journal of surfaces and colloids (2019): 7191-7204
N2 -Substituted 2'-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ.
Authors: Gowda, A S Prakasha and Lee, Marietta and Spratt, Thomas E
Journal: Angewandte Chemie (International ed. in English) (2017): 2628-2631
Fluorescent detection of copper(II) based on DNA-templated click chemistry and graphene oxide.
Authors: Zhou, Lifen and Shen, Qinpeng and Zhao, Peng and Xiang, Bingbing and Nie, Zhou and Huang, Yan and Yao, Shouzhuo
Journal: Methods (San Diego, Calif.) (2013): 299-304
Superparamagnetic iron oxide nanotheranostics for targeted cancer cell imaging and pH-dependent intracellular drug release.
Authors: Zou, Peng and Yu, Yanke and Wang, Y Andrew and Zhong, Yanqiang and Welton, Amanda and Galbán, Craig and Wang, Shaomeng and Sun, Duxin
Journal: Molecular pharmaceutics (2010): 1974-84