AAT Bioquest

FastClick™ XFD488 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 weight860.75
Spectral properties
Correction Factor (260 nm)0.30
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)71000
Excitation (nm)499
Emission (nm)520
Quantum yield0.921
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™ XFD488 Alkyne


See also: Click Chemistry
Molecular weight
Correction Factor (260 nm)
Correction Factor (280 nm)
Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
Quantum yield
FastClick™ XFD488 Azide contains both the CAG moiety of FastClick (for assisting click efficiency) and Alexa Fluor® 488 fluorophore (as the fluorescence tag) for developing Alexa Fluor® 488-based fluorescent probes. Alexa Fluor® 488 is a commonly used green fluorophore for labeling proteins, nucleic acids, or other biomolecules. It has excellent photostability and excitation that matches the common 488 nm Argon laser line that is equipped with almost all the major fluorescence instruments. Its conjugates are widely used for imaging and flow cytometry applications in combination with the widely available FITC filter set. It is a water-soluble rhodamine 110 derivative that has pH-insensitive fluorescence from pH 4 to pH 10. Alexa Fluor® is a trademark of ThermoFisher Scientific. 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™ XFD488 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 mM116.178 µL580.889 µL1.162 mL5.809 mL11.618 mL
5 mM23.236 µL116.178 µL232.356 µL1.162 mL2.324 mL
10 mM11.618 µL58.089 µL116.178 µL580.889 µL1.162 mL

Molarity calculator

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Spectral properties

Correction Factor (260 nm)0.30
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)71000
Excitation (nm)499
Emission (nm)520
Quantum yield0.921

Product Family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
FastClick™ Cy3 Azide55556915000010.1510.070.073
FastClick™ Cy5 Azide65167025000010.271, 0.420.020.03
FastClick™ Cy7 Azide7567792500000.30.050.036
FastClick™ XFD350 Azide34344119000-0.250.19
FastClick™ XFD555 Azide5535681500000.110.080.08
FastClick™ XFD647 Azide6506712390000.3310.000.03
FastClick™ XFD750 Azide7527762400000.1210.000.04



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