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AAT Bioquest

FastClick™ XFD555 Alkyne

FastClick™ XFD555 Alkyne 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. FastClick™ XFD555 Alkyne readily reacts with azido-modified biomolecules with high yields. 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.

Spectrum

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
FastClick™ Cy3 Alkyne55556915000010.1510.070.073
FastClick™ Cy5 Alkyne65167025000010.271, 0.420.020.03
FastClick™ Cy7 Alkyne7567792500000.30.050.036
FastClick™ XFD350 Alkyne34344119000-0.250.19
FastClick™ XFD488 Alkyne499520710000.9210.300.11
FastClick™ XFD647 Alkyne6506712390000.3310.000.03
FastClick™ XFD750 Alkyne7527762400000.1210.000.04

References

View all 26 references: Citation Explorer
Mechanism of Cyanine5 to Cyanine3 Photoconversion and Its Application for High-Density Single-Particle Tracking in a Living Cell.
Authors: Cho, Yoonjung and An, Hyeong Jeon and Kim, Taehoon and Lee, Chulbom and Lee, Nam Ki
Journal: Journal of the American Chemical Society (2021): 14125-14135
Molecular and Spectroscopic Characterization of Green and Red Cyanine Fluorophores from the Alexa Fluor and AF Series*.
Authors: Gebhardt, Christian and Lehmann, Martin and Reif, Maria M and Zacharias, Martin and Gemmecker, Gerd and Cordes, Thorben
Journal: Chemphyschem : a European journal of chemical physics and physical chemistry (2021)
Restricted intramolecular rotation of fluorescent molecular rotors at the periphery of aqueous microdroplets in oil.
Authors: Kang, Jooyoun and Lhee, SangMoon and Lee, Jae Kyoo and Zare, Richard N and Nam, Hong Gil
Journal: Scientific reports (2020): 16859
Real-time in vivo imaging of extracellular ATP in the brain with a hybrid-type fluorescent sensor.
Authors: Kitajima, Nami and Takikawa, Kenji and Sekiya, Hiroshi and Satoh, Kaname and Asanuma, Daisuke and Sakamoto, Hirokazu and Takahashi, Shodai and Hanaoka, Kenjiro and Urano, Yasuteru and Namiki, Shigeyuki and Iino, Masamitsu and Hirose, Kenzo
Journal: eLife (2020)
Graphene-Based Steganographic Aptasensor for Information Computing and Monitoring Toxins of Biofilm in Food.
Authors: Wang, Qi and Yang, Qingli and Wu, Wei
Journal: Frontiers in microbiology (2019): 3139
Page updated on October 28, 2024

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Catalog Number72877
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Physical properties

Molecular weight

1255.42

Solvent

DMSO

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 yield

0.11

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
The reaction (Green Bar) of FastClick Cy5 Alkyne with coumarin azide 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 alkyne does not effectively react with the coumarin azide substrate.
The reaction (Green Bar) of FastClick Cy5 Alkyne with coumarin azide 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 alkyne does not effectively react with the coumarin azide substrate.
The reaction (Green Bar) of FastClick Cy5 Alkyne with coumarin azide 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 alkyne does not effectively react with the coumarin azide substrate.
Gallery Image 2

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