AAT Bioquest

Cy3.5 tetrazine

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Physical properties
Molecular weight1377.80
Spectral properties
Correction Factor (260 nm)0.08
Correction Factor (280 nm)0.178
Extinction coefficient (cm -1 M -1)150000
Excitation (nm)579
Emission (nm)591
Quantum yield0.15
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)
Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
Quantum yield
Cy3.5 tetrazine is an excellent building block that can be readily used to label TCO-modified biological molecules for 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 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 Cy3.5 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 mM72.579 µL362.897 µL725.795 µL3.629 mL7.258 mL
5 mM14.516 µL72.579 µL145.159 µL725.795 µL1.452 mL
10 mM7.258 µL36.29 µL72.579 µL362.897 µL725.795 µL

Molarity calculator

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Spectral properties

Correction Factor (260 nm)0.08
Correction Factor (280 nm)0.178
Extinction coefficient (cm -1 M -1)150000
Excitation (nm)579
Emission (nm)591
Quantum yield0.15

Product Family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)Correction Factor (482 nm)Correction Factor (565 nm)Correction Factor (650 nm)
Cy5.5 tetrazine6837032500000.270.050.1010.00170.0470.454



View all 7 references: Citation Explorer
Evaluation of bioorthogonally applicable tetrazine-Cy3 probes for fluorogenic labeling schemes.
Authors: Albitz, Evelin and Németh, Krisztina and Knorr, Gergely and Kele, Péter
Journal: Organic & biomolecular chemistry (2023): 7358-7366
Analysis of protein phosphorylation in solution and in cells by using an ATP analogue in combination with fluorescence techniques.
Authors: Li, Yue and Huang, Xiangyi and Ren, Jicun
Journal: The Analyst (2021): 4506-4514
Design, Synthesis, Conjugation, and Reactivity of Novel trans,trans-1,5-Cyclooctadiene-Derived Bioorthogonal Linkers.
Authors: Longo, Beatrice and Zanato, Chiara and Piras, Monica and Dall'Angelo, Sergio and Windhorst, Albert D and Vugts, Danielle J and Baldassarre, Massimiliano and Zanda, Matteo
Journal: Bioconjugate chemistry (2020)
Bioorthogonally Applicable Fluorogenic Cyanine-Tetrazines for No-Wash Super-Resolution Imaging.
Authors: Knorr, Gergely and Kozma, Eszter and Schaart, Judith M and Németh, Krisztina and Török, György and Kele, Péter
Journal: Bioconjugate chemistry (2018): 1312-1318
A rapid and concise setup for the fast screening of FRET pairs using bioorthogonalized fluorescent dyes.
Authors: Petrovics, Réka and Söveges, Bianka and Egyed, Alexandra and Knorr, Gergely and Kormos, Attila and Imre, Tímea and Török, György and Zeke, András and Kocsmár, Éva and Lotz, Gábor and Kele, Péter and Németh, Krisztina
Journal: Organic & biomolecular chemistry (2018): 2997-3005
Azide and trans-cyclooctene dUTPs: incorporation into DNA probes and fluorescent click-labelling.
Authors: Ren, Xiaomei and El-Sagheer, Afaf H and Brown, Tom
Journal: The Analyst (2015): 2671-8
Rapid labeling of metabolically engineered cell-surface glycoconjugates with a carbamate-linked cyclopropene reporter.
Authors: Späte, Anne-Katrin and Bußkamp, Holger and Niederwieser, Andrea and Schart, Verena F and Marx, Andreas and Wittmann, Valentin
Journal: Bioconjugate chemistry (2014): 147-54