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AAT Bioquest

FastClick™ 6-TAMRA 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 weight642.72
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™ 6-TAMRA Alkyne

OverviewpdfSDSpdfProtocol


See also: Click Chemistry
Molecular weight
642.72
FastClick™ 6-TAMRA Azide contains both the CAG moiety of FastClick (for assisting click efficiency) and 6-TAMRA fluorophore (as the fluorescence tag) for developing 6-TAMRA-based fluorescent probes. 6-TAMRA is one of the most widely used orange fluorophores, in particular, for labeling oligonucleotides. It has almost the identical fluorescence spectra to Alexa Fluor 546, a rhodamine analog. 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™ 6-TAMRA 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 mM155.589 µL777.944 µL1.556 mL7.779 mL15.559 mL
5 mM31.118 µL155.589 µL311.177 µL1.556 mL3.112 mL
10 mM15.559 µL77.794 µL155.589 µL777.944 µL1.556 mL

Molarity calculator

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

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References


View all 7 references: Citation Explorer
Divergent Synthesis of Ultrabright and Dendritic Xanthenes for Enhanced Click-Chemistry-Based Bioimaging.
Authors: Montiel, Luis and Spada, Fabio and Crisp, Antony and Serdjukow, Sascha and Carell, Thomas and Frischmuth, Thomas
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2023): e202202633
N-Terminal selective modification of peptides and proteins using 2-ethynylbenzaldehydes.
Authors: Deng, Jie-Ren and Lai, Nathanael Chun-Him and Kung, Karen Ka-Yan and Yang, Bin and Chung, Sai-Fung and Leung, Alan Siu-Lun and Choi, Man-Chung and Leung, Yun-Chung and Wong, Man-Kin
Journal: Communications chemistry (2020): 67
Maleimide-Based Chemical Proteomics for Quantitative Analysis of Cysteine Reactivity.
Authors: McConnell, Evan W and Smythers, Amanda L and Hicks, Leslie M
Journal: Journal of the American Society for Mass Spectrometry (2020)
The application of a novel, cell permeable activity-based probe for the detection of cysteine cathepsins.
Authors: Hughes, Caroline S and Shaw, George and Burden, Roberta E and Scott, Christopher J and Gilmore, Brendan F
Journal: Biochemical and biophysical research communications (2016): 444-50
Nucleic acid sensing by an orthogonal reporter system based on homo-DNA.
Authors: Stoop, Matthias and Désiron, Camille and Leumann, Christian J
Journal: Artificial DNA, PNA & XNA (2013): 28-33
Homo-DNA templated chemistry and its application to nucleic acid sensing.
Authors: Stoop, Matthias and Leumann, Christian J
Journal: Chemical communications (Cambridge, England) (2011): 7494-6
Photo-leucine incorporation reveals the target of a cyclodepsipeptide inhibitor of cotranslational translocation.
Authors: MacKinnon, Andrew L and Garrison, Jennifer L and Hegde, Ramanujan S and Taunton, Jack
Journal: Journal of the American Chemical Society (2007): 14560-1