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AAT Bioquest

FastClick™ Biotin Azide

FastClick™ Biotin Azide contains both the CAG moiety of FastClick (for assisting click efficiency) and biotin hapten (as the detection tag) for developing biotin-based probes. It readily reacts with an alkyne-containing biomolecule under extremely mild conditions. Biotin is one of the most commonly used tags that finds a variety of utilities in biological detections similarly to other popular haptens such as 2,4-Dinitrophenol (DNP) and DIG etc. Biotin 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. 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.

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
FastClick™ XFD750 Azide7527762400000.1210.000.04

References

View all 28 references: Citation Explorer
Rapid cell type-specific nascent proteome labeling in Drosophila.
Authors: Villalobos-Cantor, Stefanny and Barrett, Ruth M and Condon, Alec F and Arreola-Bustos, Alicia and Rodriguez, Kelsie M and Cohen, Michael S and Martin, Ian
Journal: eLife (2023)
Capture, Release, and Identification of Newly Synthesized Proteins for Improved Profiling of Functional Translatomes.
Authors: Phillips, Nancy J and Vinaithirthan, Bala M and Oses-Prieto, Juan A and Chalkley, Robert J and Burlingame, Alma L
Journal: Molecular & cellular proteomics : MCP (2023): 100497
Selective Removal of Unhydrolyzed Monolinked Peptides from Enriched Crosslinked Peptides To Improve the Coverage of Protein Complex Analysis.
Authors: An, Yuxin and Zhao, Qun and Gao, Hang and Zhao, Lili and Li, Xiao and Zhang, Xiaodan and Liang, Zhen and Zhang, Lihua and Zhang, Yukui
Journal: Analytical chemistry (2022): 3904-3913
Ultrastructural localization of de novo synthesized phosphatidylcholine in yeast cells by freeze-fracture electron microscopy.
Authors: Tsuji, Takuma and Fujimoto, Toyoshi
Journal: STAR protocols (2021): 100990
Profiling and Validation of Live-Cell Protein Methylation with Engineered Enzymes and Methionine Analogues.
Authors: Weiss, Nicole and Seneviranthe, Chamara and Jiang, Ming and Wang, Ke and Luo, Minkui
Journal: Current protocols (2021): e213
Page updated on November 21, 2024

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Catalog Number72801
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Physical properties

Molecular weight

456.57

Solvent

DMSO

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
Product Image
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Gallery Image 1
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.

Alternative formats