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AAT Bioquest

FastClick™ XFD750 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 weight1211.33
SolventDMSO
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 yield0.121
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™ XFD750 Alkyne

OverviewpdfSDSpdfProtocol


See also: Click Chemistry
Molecular weight
1211.33
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
FastClick™ XFD750 Azide 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 alkyne-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.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of FastClick™ XFD750 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 mM82.554 µL412.769 µL825.539 µL4.128 mL8.255 mL
5 mM16.511 µL82.554 µL165.108 µL825.539 µL1.651 mL
10 mM8.255 µL41.277 µL82.554 µL412.769 µL825.539 µL

Molarity calculator

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Spectrum


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spectrum

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 yield0.121

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™ XFD555 Azide5535681500000.110.080.08
FastClick™ XFD647 Azide6506712390000.3310.000.03

Images


References


View all 32 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
pH-responsive graphene oxide loaded with targeted peptide and anticancer drug for OSCC therapy.
Authors: Li, Ran and Gao, Ruifang and Zhao, Yingjiao and Zhang, Fang and Wang, Xiangyu and Li, Bing and Wang, Lu and Ma, Lixin and Du, Jie
Journal: Frontiers in oncology (2022): 930920
Near-Infrared Fluorescence Imaging of Carotid Plaques in an Atherosclerotic Murine Model.
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Journal: Biomolecules (2021)
Challenging a Preconception: Optoacoustic Spectrum Differs from the Optical Absorption Spectrum of Proteins and Dyes for Molecular Imaging.
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Journal: Analytical chemistry (2020)
CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.
Authors: Kleinmanns, Katrin and Fosse, Vibeke and Davidson, Ben and de Jalón, Elvira García and Tenstad, Olav and Bjørge, Line and McCormack, Emmet
Journal: EBioMedicine (2020): 102783
Mechanistic profiling of the release kinetics of siRNA from lipidoid-polymer hybrid nanoparticles in vitro and in vivo after pulmonary administration.
Authors: Thanki, Kaushik and van Eetvelde, Delphine and Geyer, Antonia and Fraire, Juan and Hendrix, Remi and Van Eygen, Hannelore and Putteman, Emma and Sami, Haider and de Souza Carvalho-Wodarz, Cristiane and Franzyk, Henrik and Nielsen, Hanne Mørck and Braeckmans, Kevin and Lehr, Claus-Michael and Ogris, Manfred and Foged, Camilla
Journal: Journal of controlled release : official journal of the Controlled Release Society (2019): 82-93
Generation and characterization of novel recombinant anti-hERG1 scFv antibodies for cancer molecular imaging.
Authors: Duranti, Claudia and Carraresi, Laura and Sette, Angelica and Stefanini, Matteo and Lottini, Tiziano and Crescioli, Silvia and Crociani, Olivia and Iamele, Luisa and De Jonge, Hugo and Gherardi, Ermanno and Arcangeli, Annarosa
Journal: Oncotarget (2018): 34972-34989
Enhanced Release of Molecules upon Ultraviolet (UV) Light Irradiation from Photoresponsive Hydrogels Prepared from Bifunctional Azobenzene and Four-Arm Poly(ethylene glycol).
Authors: Rastogi, Shiva K and Anderson, Hailee E and Lamas, Joseph and Barret, Scott and Cantu, Travis and Zauscher, Stefan and Brittain, William J and Betancourt, Tania
Journal: ACS applied materials & interfaces (2018): 30071-30080
Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging.
Authors: Dong, Lili and Zhou, Hongxin and Zhao, Menglong and Gao, Xinghui and Liu, Yang and Liu, Dongli and Guo, Wei and Hu, Hongwei and Xie, Qian and Fan, Jia and Lin, Jiang and Wu, Weizhong
Journal: Molecular therapy. Nucleic acids (2018): 376-386
In vivo fluorescence imaging of hepatocellular carcinoma using a novel GPC3-specific aptamer probe.
Authors: Zhao, Menglong and Dong, Lili and Liu, Zhuang and Yang, Shuohui and Wu, Weizhong and Lin, Jiang
Journal: Quantitative imaging in medicine and surgery (2018): 151-160