AAT Bioquest

Cy5.5 tetrazine

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Physical properties
Molecular weight1403.84
Spectral properties
Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.101
Correction Factor (482 nm)0.0017
Correction Factor (565 nm)0.047
Correction Factor (650 nm)0.454
Extinction coefficient (cm -1 M -1)250000
Excitation (nm)683
Emission (nm)703
Quantum yield0.27
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


Molecular weight
Correction Factor (260 nm)
Correction Factor (280 nm)
Correction Factor (482 nm)
Correction Factor (565 nm)
Correction Factor (650 nm)
Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
Quantum yield
Cy5.5 tetrazine is an excellent building block that can be readily used to label TCO-modified biological molecules for NIR fluorescence imaging and other fluorescence-based biochemical analysis. It is widely used for labeling TCO-modified peptides, proteins and oligos etc. The inverse electron demand Diels–Alder (IEDDA) between 1,2,4,5-tetrazine and strained alkene (such as trans-cyclooctene, TCO) is a well-established bioorthogonal reaction. It is generally considered to be the fastest click reaction with first-order rate constants ranging up to 100,000 M−1 S−1. Since the first report on IEDDA reaction, several kinds of strained alkenes/alkynes and tetrazine analogs have been synthesized, and these functional group pairs have been applied to drug discovery, life science research and nanomaterials. Due to the extremely rapid reaction rate of IEDDA under mild conditions such as room temperature, neutral pH, and in aqueous media, this reaction has been a highly useful ligation approach for a variety of bioconjugations. AAT Bioquest offers a group of tetrazine- and TCO-containing dyes for exploring various biological systems that can use this powerful click reaction.


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Cy5.5 tetrazine 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 mM71.233 µL356.166 µL712.332 µL3.562 mL7.123 mL
5 mM14.247 µL71.233 µL142.466 µL712.332 µL1.425 mL
10 mM7.123 µL35.617 µL71.233 µL356.166 µL712.332 µL

Molarity calculator

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Spectral properties

Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.101
Correction Factor (482 nm)0.0017
Correction Factor (565 nm)0.047
Correction Factor (650 nm)0.454
Extinction coefficient (cm -1 M -1)250000
Excitation (nm)683
Emission (nm)703
Quantum yield0.27

Product Family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
Cy3.5 tetrazine5795911500000.150.080.178



View all 8 references: Citation Explorer
Optimization of IEDDA bioorthogonal system: Efficient process to improve trans-cyclooctene/tetrazine interaction.
Authors: Béquignat, Jean-Baptiste and Ty, Nancy and Rondon, Aurélie and Taiariol, Ludivine and Degoul, Françoise and Canitrot, Damien and Quintana, Mercedes and Navarro-Teulon, Isabelle and Miot-Noirault, Elisabeth and Boucheix, Claude and Chezal, Jean-Michel and Moreau, Emmanuel
Journal: European journal of medicinal chemistry (2020): 112574
Labeling a TCO-functionalized single domain antibody fragment with 18F via inverse electron demand Diels Alder cycloaddition using a fluoronicotinyl moiety-bearing tetrazine derivative.
Authors: Zhou, Zhengyuan and Zalutsky, Michael R and Vaidyanathan, Ganesan
Journal: Bioorganic & medicinal chemistry (2020): 115634
Tetrazine-TCO Ligation: A Potential Simple Approach to Improve Tumor Uptake through Enhanced Blood Circulation.
Authors: Wang, Mengzhe and Wang, Hui and Niu, Crystal Q and Zhang, Tao and Wu, Zhanhong and Li, Zibo
Journal: Bioconjugate chemistry (2020): 1795-1803
Development of a self-immolative linker for tetrazine-triggered release of alcohols in cells.
Authors: Davies, Sarah and Oliveira, Bruno L and Bernardes, Gonçalo J L
Journal: Organic & biomolecular chemistry (2019): 5725-5730
An Efficient Method for Labeling Single Domain Antibody Fragments with 18F Using Tetrazine- Trans-Cyclooctene Ligation and a Renal Brush Border Enzyme-Cleavable Linker.
Authors: Zhou, Zhengyuan and Devoogdt, Nick and Zalutsky, Michael R and Vaidyanathan, Ganesan
Journal: Bioconjugate chemistry (2018): 4090-4103
A Versatile Microarray Immobilization Strategy Based on a Biorthogonal Reaction Between Tetrazine and Trans-Cyclooctene.
Authors: Wang, Ping and Gao, Liqian and Lei, Haipeng and Lee, Su Seong and Yao, Shao Q and Sun, Hongyan
Journal: Methods in molecular biology (Clifton, N.J.) (2017): 67-80
Multiplex imaging and cellular target identification of kinase inhibitors via an affinity-based proteome profiling approach.
Authors: Su, Ying and Pan, Sijun and Li, Zhengqiu and Li, Lin and Wu, Xiaoyuan and Hao, Piliang and Sze, Siu Kwan and Yao, Shao Q
Journal: Scientific reports (2015): 7724
Microarray immobilization of biomolecules using a fast trans-cyclooctene (TCO)-tetrazine reaction.
Authors: Wang, Ping and Na, Zhenkun and Fu, Jiaqi and Tan, Chelsea Y J and Zhang, Huatang and Yao, Shao Q and Sun, Hongyan
Journal: Chemical communications (Cambridge, England) (2014): 11818-21