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AAT Bioquest

FastClick™ XFD555 Azide

FastClick™ XFD555 Azide contains both the CAG moiety of FastClick (for assisting click efficiency) and Alexa Fluor® 555 fluorophore (as the fluorescence tag) for developing Alexa Fluor® 555-based fluorescent probes. Alexa Fluor® 555 is a commonly used orange-red fluorophore for labeling proteins, nucleic acids, or other biomolecules. It has moderate photostability and excitation that matches the common 555 nm laser line that is equipped with almost most of the major fluorescence instruments. Its conjugates are widely used for imaging and flow cytometry applications in combination with the widely available Cy3 filter set. It is a water-soluble Cy3 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.
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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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Catalog Number72737
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Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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Physical properties
Molecular weight1159.25
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.08
Correction Factor (280 nm)0.08
Extinction coefficient (cm -1 M -1)150000
Excitation (nm)553
Emission (nm)568
Quantum yield0.11
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
Spectrum
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™ XFD488 Azide499520710000.9210.300.11
FastClick™ XFD647 Azide6506712390000.3310.000.03
FastClick™ XFD750 Azide7527762400000.1210.000.04
References
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Journal: Biosensors & bioelectronics (2023): 115339
Fluorescence Intensity and Fluorescence Lifetime Imaging Microscopies (FLIM) of Cell Differentiation in the Small Intestinal Organoids Using Cholera Toxin.
Authors: Okkelman, Irina A and Dmitriev, Ruslan I
Journal: Methods in molecular biology (Clifton, N.J.) (2023): 171-195
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Journal: Biomedical optics express (2023): 550-558