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AAT Bioquest

FastClick™ XFD488 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 weight956.92
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.30
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)71000
Excitation (nm)499
Emission (nm)520
Quantum yield0.921
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™ XFD488 Azide

OverviewpdfSDSpdfProtocol


See also: Click Chemistry
Molecular weight
956.92
Correction Factor (260 nm)
0.30
Correction Factor (280 nm)
0.11
Extinction coefficient (cm -1 M -1)
71000
Excitation (nm)
499
Emission (nm)
520
Quantum yield
0.921
FastClick™ XFD488 Alkyne contains both the CAG moiety of FastClick (for assisting click efficiency) and Alexa Fluor® 488 fluorophore (as the fluorescence tag) for developing Alexa Fluor® 488-based fluorescent probes. FastClick™ XFD405 Alkyne readily reacts with azido-modified biomolecules with high yields. Alexa Fluor® 488 is a predominantly used green fluorophore for labeling proteins, nucleic acids, or other biomolecules. It has excellent photostability and excitation that matches the common 488 nm Argon laser line that is equipped with almost all the major fluorescence instruments. Its conjugates are widely used for imaging and flow cytometry applications in combination with the widely available FITC filter set. It is a water-soluble rhodamine 110 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.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of FastClick™ XFD488 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 mM104.502 µL522.51 µL1.045 mL5.225 mL10.45 mL
5 mM20.9 µL104.502 µL209.004 µL1.045 mL2.09 mL
10 mM10.45 µL52.251 µL104.502 µL522.51 µL1.045 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)0.30
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)71000
Excitation (nm)499
Emission (nm)520
Quantum yield0.921

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™ XFD555 Alkyne5535681500000.110.080.08
FastClick™ XFD647 Alkyne6506712390000.3310.000.03
FastClick™ XFD750 Alkyne7527762400000.1210.000.04

Images


References


View all 24 references: Citation Explorer
Recyclable cell-surface chemical tags for repetitive cancer targeting.
Authors: Bhatta, Rimsha and Han, Joonsu and Zhou, Jingyi and Li, Haoyu and Wang, Hua
Journal: Journal of controlled release : official journal of the Controlled Release Society (2022): 164-174
Clickable Galactose Analogues for Imaging Glycans in Developing Zebrafish.
Authors: Daughtry, Jessica L and Cao, Wendy and Ye, Johnny and Baskin, Jeremy M
Journal: ACS chemical biology (2020): 318-324
Nanobody click chemistry for convenient site-specific fluorescent labelling, single step immunocytochemistry and delivery into living cells by photoporation and live cell imaging.
Authors: Hebbrecht, Tim and Liu, Jing and Zwaenepoel, Olivier and Boddin, Gaëlle and Van Leene, Chloé and Decoene, Klaas and Madder, Annemieke and Braeckmans, Kevin and Gettemans, Jan
Journal: New biotechnology (2020): 33-43
Tylophorine reduces protein biosynthesis and rapidly decreases cyclin D1, inhibiting vascular smooth muscle cell proliferation in vitro and in organ culture.
Authors: Joa, Helge and Blažević, Tina and Grojer, Christoph and Zeller, Iris and Heiss, Elke H and Atanasov, Atanas G and Feldler, Ines and Gruzdaitis, Päivi and Czaloun, Christa and Proksch, Peter and Messner, Barbara and Bernhard, David and Dirsch, Verena M
Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology (2019): 152938
PET imaging of EGFR expression using an 18F-labeled RNA aptamer.
Authors: Cheng, Siyuan and Jacobson, Orit and Zhu, Guizhi and Chen, Zhen and Liang, Steve H and Tian, Rui and Yang, Zhen and Niu, Gang and Zhu, Xiaohua and Chen, Xiaoyuan
Journal: European journal of nuclear medicine and molecular imaging (2019): 948-956
Detection of oriC-Independent Replication in Escherichia coli Cells.
Authors: Martel, Makisha and Balleydier, Aurélien and Brochu, Julien and Drolet, Marc
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 131-138
Flow cytometry in formamide treated cells.
Authors: Radicchio, Giovanna and Colicchia, Valeria and Marrapodi, Ramona and Carbonari, Maurizio
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2018): 829-836
Intracellular in situ labeling of TiO2 nanoparticles for fluorescence microscopy detection.
Authors: Brown, Koshonna and Thurn, Ted and Xin, Lun and Liu, William and Bazak, Remon and Chen, Si and Lai, Barry and Vogt, Stefan and Jacobsen, Chris and Paunesku, Tatjana and Woloschak, Gayle E
Journal: Nano research (2018): 464-476
Fluorescent Labeling of Collagen Production by Cells for Noninvasive Imaging of Extracellular Matrix Deposition.
Authors: Bardsley, Katie and Yang, Ying and El Haj, Alicia J
Journal: Tissue engineering. Part C, Methods (2017): 228-236
Isolation of Plant Nuclei at Defined Cell Cycle Stages Using EdU Labeling and Flow Cytometry.
Authors: Wear, Emily E and Concia, Lorenzo and Brooks, Ashley M and Markham, Emily A and Lee, Tae-Jin and Allen, George C and Thompson, William F and Hanley-Bowdoin, Linda
Journal: Methods in molecular biology (Clifton, N.J.) (2016): 69-86