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AAT Bioquest

FastClick™ Cy7 Alkyne

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.
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Physical properties
Molecular weight991.28
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.036
Correction Factor (482 nm)0.0005
Correction Factor (565 nm)0.0193
Correction Factor (650 nm)0.165
Extinction coefficient (cm -1 M -1)250000
Excitation (nm)756
Emission (nm)779
Quantum yield0.3
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™ Cy7 Azide

OverviewpdfSDSpdfProtocol


See also: Click Chemistry
Molecular weight
991.28
Correction Factor (260 nm)
0.05
Correction Factor (280 nm)
0.036
Correction Factor (482 nm)
0.0005
Correction Factor (565 nm)
0.0193
Correction Factor (650 nm)
0.165
Extinction coefficient (cm -1 M -1)
250000
Excitation (nm)
756
Emission (nm)
779
Quantum yield
0.3
FastClick™ Cy7 Alkyne contains both the moiety of FastClick (for assisting click efficiency) and Cy7 fluorophore (as the fluorescence tag) for developing Cy7-based fluorescent probes. It readily reacts with an azido-modified biomolecule under extremely mild conditions. Cy7 is one of the most common near infrared (NIR) fluorophores used for developing in vivo imaging probes. Cy7 antibody conjugates are widely used in fluorescence imaging and flow cytometric applications. 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.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of FastClick™ Cy7 Alkyne 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 mM100.88 µL504.398 µL1.009 mL5.044 mL10.088 mL
5 mM20.176 µL100.88 µL201.759 µL1.009 mL2.018 mL
10 mM10.088 µL50.44 µL100.88 µL504.398 µL1.009 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


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spectrum

Spectral properties

Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.036
Correction Factor (482 nm)0.0005
Correction Factor (565 nm)0.0193
Correction Factor (650 nm)0.165
Extinction coefficient (cm -1 M -1)250000
Excitation (nm)756
Emission (nm)779
Quantum yield0.3

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™ XFD350 Alkyne34344119000-0.250.19
FastClick™ XFD488 Alkyne499520710000.9210.300.11
FastClick™ XFD555 Alkyne5535681500000.110.080.08
FastClick™ XFD647 Alkyne6506712390000.3310.000.03
FastClick™ XFD750 Alkyne7527762400000.1210.000.04

Images


References


View all 13 references: Citation Explorer
Loading Intracranial Drug-Eluting Reservoirs Across the Blood-Brain Barrier With Focused Ultrasound.
Authors: Moody, Christopher T and Durham, Phillip G and Dayton, Paul A and Brudno, Yevgeny
Journal: Ultrasound in medicine & biology (2023): 1679-1685
Quantitative and biosafe modification of bifunctional groups onto carbon dots by click chemistry.
Authors: He, Qian and Wu, Zewen and Li, Jingxuan and Li, Ruijiao and Zhang, Liyun and Liu, Yaodong
Journal: Journal of materials chemistry. B (2023): 5094-5100
Differential Translation Activity Analysis Using Bioorthogonal Noncanonical Amino Acid Tagging (BONCAT) in Archaea.
Authors: Kern, Michael and Ferreira-Cerca, Sébastien
Journal: Methods in molecular biology (Clifton, N.J.) (2022): 229-246
Controlling Coagulation in Blood with Red Light.
Authors: Müller, Patricia and Sahlbach, Marlen and Gasper, Simone and Mayer, Günter and Müller, Jens and Pötzsch, Bernd and Heckel, Alexander
Journal: Angewandte Chemie (International ed. in English) (2021): 22441-22446
Clickable Methyltetrazine-Indocarbocyanine Lipids: A Multicolor Tool Kit for Efficient Modifications of Cell Membranes.
Authors: Gaikwad, Hanmant and Wang, Guankui and Smith, Weston J and Alexander, Keisha L and D'Alessandro, Angelo and Zhang, Wei and Purev, Enkhtsetseg and Simberg, Dmitri
Journal: Bioconjugate chemistry (2019): 2106-2114
Erlotinib-Guided Self-Assembled Trifunctional Click Nanotheranostics for Distinguishing Druggable Mutations and Synergistic Therapy of Nonsmall Cell Lung Cancer.
Authors: Gao, Yu and Zhang, Huijuan and Zhang, Yingying and Lv, Tingting and Zhang, Lu and Li, Ziying and Xie, Xiaodong and Li, Fengqiao and Chen, Haijun and Jia, Lee
Journal: Molecular pharmaceutics (2018): 5146-5161
Bioorthogonally Applicable Fluorescence Deactivation Strategy for Receptor Kinetics Study and Theranostic Pretargeting Approaches.
Authors: van der Wal, Steffen and de Korne, Clarize M and Sand, Laurens G L and van Willigen, Danny M and Hogendoorn, Pancras C W and Szuhai, Karoly and van Leeuwen, Fijs W B and Buckle, Tessa
Journal: Chembiochem : a European journal of chemical biology (2018)
Folate and Heptamethine Cyanine Modified Chitosan-Based Nanotheranostics for Tumor Targeted Near-Infrared Fluorescence Imaging and Photodynamic Therapy.
Authors: Zhang, Yingying and Lv, Tingting and Zhang, Huijuan and Xie, Xiaodong and Li, Ziying and Chen, Haijun and Gao, Yu
Journal: Biomacromolecules (2017): 2146-2160
Flow cytometric analysis of T lymphocyte proliferation in vivo by EdU incorporation.
Authors: Sun, Xiaojing and Zhang, Chunpan and Jin, Hua and Sun, Guangyong and Tian, Yue and Shi, Wen and Zhang, Dong
Journal: International immunopharmacology (2016): 56-65
NIR-Cyanine Dye Linker: a Promising Candidate for Isochronic Fluorescence Imaging in Molecular Cancer Diagnostics and Therapy Monitoring.
Authors: Komljenovic, Dorde and Wiessler, Manfred and Waldeck, Waldemar and Ehemann, Volker and Pipkorn, Ruediger and Schrenk, Hans-Hermann and Debus, Jürgen and Braun, Klaus
Journal: Theranostics (2016): 131-41