iFluor® 555 TCO

The tetrazine-trans-cyclooctene (TCO) ligation constitutes a non-toxic biomolecule labeling method of unparalleled speed. A tetrazine-functionalized molecule reacts with a TCO-functionalized molecule, forming a stable conjugate via a dihydropyrazine moiety. This has gained popularity due to its extremely fast kinetics. iFluor® 555 TCO can be readily used to label tetrazine-modified biological molecules for fluorescence imaging and other fluorescence-based biochemical analysis.

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	1236.49
Solvent	DMSO

Spectral properties

Correction Factor (260 nm)	0.23
Correction Factor (280 nm)	0.14
Extinction coefficient (cm ^-1 M ^-1)	100000¹
Excitation (nm)	557
Emission (nm)	570
Quantum yield	0.64¹

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

Molecular weight

1236.49

Correction Factor (260 nm)

0.23

Correction Factor (280 nm)

0.14

Extinction coefficient (cm ^-1 M ^-1)

100000¹

Excitation (nm)

557

Emission (nm)

570

Quantum yield

0.64¹

The tetrazine-trans-cyclooctene (TCO) ligation constitutes a non-toxic biomolecule labeling method of unparalleled speed. A tetrazine-functionalized molecule reacts with a TCO-functionalized molecule, forming a stable conjugate via a dihydropyrazine moiety. This has gained popularity due to its extremely fast kinetics. AAT Bioquest offers a group of tetrazine- and TCO-containing dyes for exploring various biological systems that can use this powerful click reaction. iFluor® 555-TCO can be readily used to label tetrazine-modified biological molecules for fluorescence imaging and other fluorescence-based biological applications. The peptide and oligo conjugates prepared with iFluor® 555 dye are far superior to the conjugates of other spectrally similar dyes, such as the popular Cy3 and Alexa Fluor® 555. iFluor® 555 conjugates are significantly brighter and much more photostable than the corresponding conjugates of Cy3 and Alexa Fluor® 555 (Alexa Fluor® is the trademark of Invitrogen).

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 555 TCO to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	80.874 µL	404.37 µL	808.741 µL	4.044 mL	8.087 mL
5 mM	16.175 µL	80.874 µL	161.748 µL	808.741 µL	1.617 mL
10 mM	8.087 µL	40.437 µL	80.874 µL	404.37 µL	808.741 µL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
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Spectrum

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Spectral properties

Correction Factor (260 nm)	0.23
Correction Factor (280 nm)	0.14
Extinction coefficient (cm ^-1 M ^-1)	100000¹
Excitation (nm)	557
Emission (nm)	570
Quantum yield	0.64¹

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (260 nm)	Correction Factor (280 nm)
iFluor® 555 maleimide	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 555 amine	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 555 hydrazide	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 555 alkyne	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 555 azide	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 555 Styramide Superior Replacement for Alexa Fluor 555 tyramide and Opal 570	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 555 Tyramide	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 488 TCO	491	516	75000¹	0.9¹	0.21	0.11
iFluor® 594 TCO	588	604	180000¹	0.53¹	0.05	0.04
iFluor® 647 TCO	656	670	250000¹	0.25¹	0.03	0.03
iFluor® 555 Tetrazine	557	570	100000¹	0.64¹	0.23	0.14
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Images

Figure 1. The tetrazine-trans-cyclooctene (TCO) ligation constitutes a non-toxic biomolecule labeling method of unparalleled speed. A tetrazine-functionalized molecule reacts with a TCO-functionalized molecule, forming a stable conjugate via a dihydropyrazine moiety. This has gained popularity due to its extremely fast kinetics. iFluor® 555 TCO can be readily used to label tetrazine-modified biological molecules for fluorescence imaging and other fluorescence-based biochemical analysis.

References

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Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2020)

Fluorescent bioorthogonal labeling of class B GPCRs in live cells.
Authors: Gangam, Srikanth Kumar and Lin, Qing
Journal: Methods in enzymology (2020): 95-111

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Authors: Wu, Xunshen and Wu, Kui and Gaye, Fatima and Royzen, Maksim
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Fluorescent Janus emulsions for biosensing of Listeria monocytogenes.
Authors: Li, Jie and Savagatrup, Suchol and Nelson, Zachary and Yoshinaga, Kosuke and Swager, Timothy M
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Conjugable A3 adenosine receptor antagonists for the development of functionalized ligands and their use in fluorescent probes.
Authors: Federico, Stephanie and Margiotta, Enrico and Moro, Stefano and Kozma, Eszter and Gao, Zhan-Guo and Jacobson, Kenneth A and Spalluto, Giampiero
Journal: European journal of medicinal chemistry (2020): 111886

Metabolic phospholipid labeling of intact bacteria enables a fluorescence assay that detects compromised outer membranes.
Authors: Nilsson, Inga and Lee, Sheng Y and Sawyer, William S and Baxter Rath, Christopher M and Lapointe, Guillaume and Six, David A
Journal: Journal of lipid research (2020): 870-883

Optimizing the Biodistribution of Radiofluorinated Barbiturate Tracers for Matrix Metalloproteinase Imaging by Introduction of Fluorescent Dyes as Pharmacokinetic Modulators.
Authors: Schwegmann, Katrin and Hohn, Michael and Hermann, Sven and Schäfers, Michael and Riemann, Burkhard and Haufe, Günter and Wagner, Stefan and Breyholz, Hans-Jörg
Journal: Bioconjugate chemistry (2020): 1117-1132

Synthesis and Characterization of Heterodimers and Fluorescent Nisin Species by Incorporation of Methionine Analogs and Subsequent Click Chemistry.
Authors: Deng, Jingjing and Viel, Jakob H and Chen, Jingqi and Kuipers, Oscar P
Journal: ACS synthetic biology (2020)

Ultra-fast Cycling for Multiplexed Cellular Fluorescence Imaging.
Authors: Ko, Jina and Oh, Juhyun and Ahmed, Maaz S and Carlson, Jonathan C T and Weissleder, Ralph
Journal: Angewandte Chemie (International ed. in English) (2020): 6839-6846

Color-Tunable Light-up Bioorthogonal Probes for In Vivo Two-Photon Fluorescence Imaging.
Authors: Dou, Yandong and Wang, Yajun and Duan, Yukun and Liu, Bin and Hu, Qinglian and Shen, Wei and Sun, Hongyan and Zhu, Qing
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2020): 4576-4582