Tetrazine-DBCO Data Sheet - KeraFAST

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Tetrazine‐DBCO, Triethylammonium Salt
Product No.: 1022
Product Name:
Tetrazine‐DBCO, Triethylammonium Salt
Chemical Structure:
N
N
N N
N
H
O
N
H
S
O O O
Et 3 NH
O
O
N
Chemical Composition:
Molecular Weight:
Solubility:
Storage:
Shelf life:
C 32 H 29 N 7 O 6 S (protonated)
639.68 (protonated)
Water, DMSO, DMF, MeOH
Upon receipt store at -20°C. Product shipped at ambient temperature
At least 12 month at -20 0 C
Introduction
Tetrazine‐DBCO is a water‐soluble reagent for converting of azido‐modified peptides or biopolymers into
tetrazine‐modified peptides or biopolymers. When applied in stoichiometric amount tetrazine‐DBCO can serve
as a heterobifunctional azide‐to‐TCO crosslinker for covalently linking azido‐ and TCO‐modified peptides or
biopolymers. Once a peptide or biopolymer is tetrazine‐labeled, it can be reacted with a TCO‐labeled molecule
to produce a stable conjugate (Figure 1).
Figure 1. Schematic representation of TCO-tetrazine ligation reaction.

Applications
Protein‐protein conjugation
Protein‐small molecule conjugation
18 F radiolabeling
Protein‐peptide conjugation
Surface modification
Protein‐oligonucleotide conjugation
Important Product Information
Molecules to be reacted with Tetrazine‐DBCO must have available azides.
Do not use buffers that contain azides.
Procedure for Azide-to-Tetrazine Conversion
1. Prepare the azido‐containing sample in reaction buffer or organic solvent.
2. Dissolve Tetrazine‐DBCO in aqueous buffer or organic solvent.
3. Add 2‐10 fold excess of Tetrazine‐DBCO to azido‐containing sample.
4. Incubate the reaction at room temperature for 1‐4 hour. Incubation at 4 0 C requires 2‐12 hours.
5. Remove the excess reagent by desalting the labeled protein through a desalt spin column or by azidemodified
agarose.
Procedure for TCO-Tetrazine Conjugation
1. Calculate 1.1‐5 fold molar excess of tetrazine‐ or TCO‐modified biopolymer over the corresponding
complimentary reagent.
2. Mix calculated amount of tetrazine‐labeled biopolymer with desired amount of TCO‐modified
biopolymer.
3. Allow reaction to proceed for 30 minutes at room temperature. Longer reaction time might improve
conjugation efficiency.
4. Store conjugate at 4 0 C until ready for purification or use.
References
1. Karver, M. R., et. al. (2012). "Bioorthogonal Reaction Pairs Enable Simultaneous, Selective, Multi-Target Imaging." Angew. Chem. Int.
Ed., 51:920-922.
2. Blackman, M. L., et. al. (2008). "Tetrazine Ligation: Fast Bioconjugation Based on Inverse-Electron-Demand Diels-Alder Reactivity." J.
Am. Chem. Soc., 130:13518-13519.
3. Devaraj, N. K., et. al.(2008) "Tetrazine-Based Cycloadditions: Application to Pretargeted Live Cell Imaging." Bioconjugate Chem.,
19:2297–2299.
4. Devaraj, N. K., et. al. (2009) "Fast and Sensitive Pre-Targeted Labeling of Cancer Cells through a Tetrazine/trans-Cyclooctene
Cycloaddition." Angew. Chem. Int. Ed., 48:7013-7016
5. Haun, J.B., et. al. (2009)" Probing Intracellular Biomarkers and Mediators of Cell Activation Using Nanosensor and Bioorthogonal
Chemistry" ACS Nano., 5:3204-321
For research use only
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