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AAT Bioquest

FastClick™ Biotin 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 weight552.74
SolventDMSO
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™ Biotin Azide
Related products
FastClick™ Cy3 Azide
FastClick™ Cy5 Azide
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OverviewpdfSDSpdfProtocol


See also: Click Chemistry
Molecular weight
552.74
FastClick™ Biotin Alkyne contains both the CAG moiety of FastClick (for assisting click efficiency) and biotin (as the detection tag) for developing biotin-based probes. It readily reacts with an azido-containing biomolecule under extremely mild conditions. Biotin is one of the most commonly used tags that finds a variety of utilities in biological detections. Its conjugates are widely used in fluorescence imaging, fluorescence in situ hybridization (FISH) and nucleic acid detections in combination with the fluorescence-labeled streptavidin, avidin or NeutrAvidin® or other biotin-binding protein conjugates. AAT Bioquest offers the largest collection of biotin and streptavidin products for immunological detections. 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™ Biotin 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 mM180.917 µL904.584 µL1.809 mL9.046 mL18.092 mL
5 mM36.183 µL180.917 µL361.834 µL1.809 mL3.618 mL
10 mM18.092 µL90.458 µL180.917 µL904.584 µL1.809 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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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
FastClick™ XFD750 Alkyne7527762400000.1210.000.04

Images


References


View all 16 references: Citation Explorer
Bioorthogonal Chemical Labeling Probes Targeting Sialic Acid Isomers for N-Glycan MALDI Imaging Mass Spectrometry of Tissues, Cells, and Biofluids.
Authors: Lu, Xiaowei and McDowell, Colin T and Blaschke, Calvin R K and Liu, Liping and Grimsley, Grace and Wisniewski, Luke and Gao, ChongFeng and Mehta, Anand S and Haab, Brian B and Angel, Peggi M and Drake, Richard R
Journal: Analytical chemistry (2023): 7475-7486
Tagging and Capture of Prenylated CaaX-Proteins from Plant Cell Cultures.
Authors: Ribeiro, Iliana and Ducos, Eric and Giglioli-Guivarc'h, Nathalie and Dutilleul, Christelle
Journal: Methods in molecular biology (Clifton, N.J.) (2022): 241-248
Quantitative BONCAT Allows Identification of Newly Synthesized Proteins after Optic Nerve Injury.
Authors: Shah, Sahil H and Schiapparelli, Lucio M and Yokota, Satoshi and Ma, Yuanhui and Xia, Xin and Shankar, Sahana and Saturday, Sarah and Nahmou, Michael and Sun, Catalina and Yates, John and Cline, Hollis T and Goldberg, Jeffrey L
Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience (2022): 4042-4052
Dynamics of Non-Canonical Amino Acid-Labeled Intra- and Extracellular Proteins in the Developing Mouse.
Authors: Saleh, Aya M and Jacobson, Kathryn R and Kinzer-Ursem, Tamara L and Calve, Sarah
Journal: Cellular and molecular bioengineering (2019): 495-509
Novel biotin linker with alkyne and amino groups for chemical labelling of a target protein of a bioactive small molecule.
Authors: Anabuki, Tomoaki and Tsukahara, Miu and Okamoto, Masanori and Matsuura, Hideyuki and Takahashi, Kosaku
Journal: Bioorganic & medicinal chemistry letters (2018): 783-786
Multiwalled carbon nanotubes for drug delivery: Efficiency related to length and incubation time.
Authors: Sciortino, Niccolò and Fedeli, Stefano and Paoli, Paolo and Brandi, Alberto and Chiarugi, Paola and Severi, Mirko and Cicchi, Stefano
Journal: International journal of pharmaceutics (2017): 69-72
ASG2 is a farnesylated DWD protein that acts as ABA negative regulator in Arabidopsis.
Authors: Dutilleul, Christelle and Ribeiro, Iliana and Blanc, Nathalie and Nezames, Cynthia D and Deng, Xing Wang and Zglobicki, Piotr and Palacio Barrera, Ana María and Atehortùa, Lucia and Courtois, Martine and Labas, Valérie and Giglioli-Guivarc'h, Nathalie and Ducos, Eric
Journal: Plant, cell & environment (2016): 185-98
Tandem photoaffinity labeling of a target protein using a linker with biotin, alkyne and benzophenone groups and a bioactive small molecule with an azide group.
Authors: Anabuki, Tomoaki and Tsukahara, Miu and Matsuura, Hideyuki and Takahashi, Kosaku
Journal: Bioscience, biotechnology, and biochemistry (2016): 432-9
Proteome-Wide Profiling of Targets of Cysteine reactive Small Molecules by Using Ethynyl Benziodoxolone Reagents.
Authors: Abegg, Daniel and Frei, Reto and Cerato, Luca and Prasad Hari, Durga and Wang, Chao and Waser, Jerome and Adibekian, Alexander
Journal: Angewandte Chemie (International ed. in English) (2015): 10852-7
Metabolic synthesis of clickable glutathione for chemoselective detection of glutathionylation.
Authors: Samarasinghe, Kusal T G and Munkanatta Godage, Dhanushka N P and VanHecke, Garrett C and Ahn, Young-Hoon
Journal: Journal of the American Chemical Society (2014): 11566-9