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AAT Bioquest

FastClick™ XFD750 Alkyne

FastClick™ XFD750 Alkyne contains both the CAG moiety of FastClick (for assisting click efficiency) and Alexa Fluor® 750 fluorophore (as the fluorescence tag) for developing Alexa Fluor® 750-based fluorescent probes. It readily reacts with an azido-containing biomolecule under extremely mild conditions. Alexa Fluor® 750 is a commonly used near infrared (NIR) fluorophore for labeling proteins, nucleic acids, or other biomolecules. It has moderate photostability and excitation that matches the common 750 nm laser line. Its conjugates are widely used for in vivo imaging and flow cytometry applications in combination with the widely available Cy7 filter set. It is a water-soluble Cy75 derivative that has pH-insensitive fluorescence from pH 4 to pH 10. Alexa Fluor® 750 dye is particularly useful in stochastic optical reconstruction microscopy (STORM), where it is an exceptional reporter for both dSTORM and nSTORM. It is also used in super-resolution microscopy (SRM) applications. 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™ XFD555 Alkyne5535681500000.110.080.08
FastClick™ XFD647 Alkyne6506712390000.3310.000.03

References

View all 5 references: Citation Explorer
Targeting lung cancer with clinically relevant EGFR mutations using anti-EGFR RNA aptamer.
Authors: Thomas, Brian J and Guldenpfennig, Caitlyn and Guan, Yue and Winkler, Calvin and Beecher, Margaret and Beedy, Michaela and Berendzen, Ashley F and Ma, Lixin and Daniels, Mark A and Burke, Donald H and Porciani, David
Journal: Molecular therapy. Nucleic acids (2023): 102046
Circulating Regulatory T Cell Subsets in Patients with Sarcoidosis.
Authors: Kudryavtsev, Igor and Zinchenko, Yulia and Starshinova, Anna and Serebriakova, Maria and Malkova, Anna and Akisheva, Tatiana and Kudlay, Dmitriy and Glushkova, Anzhela and Yablonskiy, Piotr and Shoenfeld, Yehuda
Journal: Diagnostics (Basel, Switzerland) (2023)
iCoreDrop: A robust immune monitoring spectral cytometry assay with six open channels for biomarker flexibility.
Authors: Jensen, Holly A and Kim, Jeong
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2022)
[Changes of monocyte and monocyte-platelet aggregates in different subgroups of thrombotic events in patients with acute myocardial infarction during PCI].
Authors: Wang, Sheng and Sun, Cuifang and Liao, Wang and Wu, Zhongwei and Wang, Yudai and Huang, Xiuxian and Lu, Sijia and Dong, Xiaoli and Shuai, Fujie and Li, Bin
Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (2017): 959-965
Quantitative comparison of long-wavelength Alexa Fluor dyes to Cy dyes: fluorescence of the dyes and their bioconjugates.
Authors: Berlier, Judith E and Rothe, Anca and Buller, Gayle and Bradford, Jolene and Gray, Diane R and Filanoski, Brian J and Telford, William G and Yue, Stephen and Liu, Jixiang and Cheung, Ching-Ying and Chang, Wesley and Hirsch, James D and Beechem, Joseph M and Haugland, Rosaria P and Haugland, Richard P
Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (2003): 1699-712
Page updated on October 11, 2024

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Catalog Number72885
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Physical properties

Molecular weight

1307.49

Solvent

DMSO

Spectral properties

Correction Factor (260 nm)

0.00

Correction Factor (280 nm)

0.04

Extinction coefficient (cm -1 M -1)

240000

Excitation (nm)

752

Emission (nm)

776

Quantum yield

0.121

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

Alternative formats