TBTA (Synonyms: Tris(benzyltriazolylmethyl)amine) |
| Catalog No.GC45003 |
TBTA (tris(benzyltriazolylmethyl)amine) is a tertiary amine containing a 1,2,3-triazole group. TBTA complexes with copper(I) and promotes click reaction catalysis by stabilizing the copper(I) oxidation state.
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Cas No.: 510758-28-8
Sample solution is provided at 25 µL, 10mM.
TBTA (tris(benzyltriazolylmethyl)amine) is a tertiary amine containing a 1,2,3-triazole group[1]. TBTA complexes with copper(I) and promotes click reaction catalysis by stabilizing the copper(I) oxidation state[2]. TBTA is one of the most widely used water-insoluble ligands in copper-catalyzed azide-alkyne cycloaddition (CuAAC)[3].
Click chemistry is a versatile reaction that can be used for the synthesis of a variety of conjugates. Virtually any biomolecule can be easily labeled with small molecules, such as fluorescent dyes, biotin, etc using the click chemistry method.
Click chemistry reaction takes place between two components: azide and alkyne (terminal acetylene). Both azido and alkyne groups are nearly never encountered in natural biomolecules. Hence, the reaction is highly bioorthogonal and specific.
References:
[1] Alireza Movahedi, et al. One-pot synthesis of TBTA-functionalized coordinating polymers. Reactive and Functional Polymers.2014. 82:1-8.
[2] Kennedy DC, McKay C S, Legault M C B, et al. Cellular consequences of copper complexes used to catalyze bioorthogonal click reactions[J]. Journal of the American Chemical Society, 2011, 133(44): 17993-18001.
[3] Hagmann V, Sommer S, Fabian P, et al. Chemically monoubiquitinated PEX5 binds to the components of the peroxisomal docking and export machinery[J]. Scientific Reports, 2018, 8(1): 16014.
This plan only provides a guide, please modify it to meet your specific needs.
1. Calculate the volumes of reagents required for Click chemistry labeling using the table below. Prepare the required stock solutions(see Appendix).
|
Reagent |
Final concentration in the mixture |
Stock solution concentration |
|
Oligonucleotide, alkyne-modified |
Varies (20 — 200 uM) |
Varies |
|
Azide |
1.5 x (oligonucleotide concentration) |
10 mM in DMSO |
|
DMSO |
50 vol % |
— |
|
Ascorbic acid |
0.5 mM |
5 mM in water |
|
Cu-TBTA complex |
0.5 mM |
10 mM in 55 vol % DMSO |
2. Dissolve alkyne-modified oligonucleotide or DNA in water in a pressure-tight vial.
3. Add 2M triethylammonium acetate buffer, pH 7.0, to final concentration 0.2 M.
4. Add DMSO, and vortex.
5. Add azide stock solution (10 mM in DMSO), and vortex.
6. Add the required volume of 5mM Ascorbic Acid Stock solution to the mixture, and vortex briefly.
7. Degass the solution by bubbling inert gas in it for 30 seconds. Nitrogen, argon, or helium can be used.
8. Add the required amount of 10 mM Copper (II)-TBTA Stock in 55% DMSO to the mixture. Flush the vial with inert gas and close the cap.
9. Vortex the mixture thoroughly. If significant precipitation of azide is observed, heat the vial for 3 minutes at 80°C, and vortex.
10. Keep at room temperature overnight.
11. Precipitate the conjugate with acetone (for oligonucleotides) or with ethanol (for DNA). Mix thoroughly and keep at −20 °C for 20 minutes.
To precipitate an oligonucleotide conjugate: add to the mixture at least a 4-fold excess volume of 3% lithium perchlorate in acetone (if the volume of the mixture is large, split in several vials).
To precipitate a DNA conjugate: add to the mixture sodium acetate to a final concentration of 0.3M; add 2.5 volumes of ethanol (or 0.8 volumes of isopropanol).
12. Centrifuge at 10000 rpm for 10 minutes.
13. Discard the supernatant. Wash the pellet with acetone (1 mL), centrifuge at 10000 rpm for 10 minutes.
14. Discard the supernatant, dry the pellet, and purify the conjugate by RP-HPLC or PAGE.
Appendix. Preparation of stock solutions of the reagents used for click-chemistry labeling and conjugation.
|
5 mM Ascorbic Acid Stock |
|
|
Preparation |
Dissolve 18 mg of ascorbic acid in 20 mL of distilled water |
|
Storage |
Ascorbic acid is readily oxidized by air. The solution is stable for one day. Use fresh preparations |
|
10 mM Copper (II)-TBTA Stock in 55% DMSO |
|
|
Preparation |
Dissolve 50 mg of copper (II) sulfate pentahydrate in 10 mL of distilled water. Dissolve 116 mg of TBTA ligand in 11 mL of DMSO. Mix two solutions |
|
Storage |
Store at room temperature. The solution is stable for years |
|
2M Triethylammonium Acetate Buffer, pH 7.0 |
|
|
Preparation |
Mix 2.78 mL of triethylamine with 1.14 mL of acetic acid. Add water to 10 mL volume, and adjust pH to 7.0 |
|
Storage |
Store at room temperature. The solution is stable for years |
| Cas No. | 510758-28-8 | SDF | |
| Synonyms | Tris(benzyltriazolylmethyl)amine | ||
| Canonical SMILES | C1(CN(CC2=CN(CC3=CC=CC=C3)N=N2)CC4=CN(CC5=CC=CC=C5)N=N4)=CN(CC6=CC=CC=C6)N=N1 | ||
| Formula | C30H30N10 | M.Wt | 530.6 |
| Solubility | DMF: 30 mg/ml,DMSO: 30 mg/ml,DMSO:PBS(pH7.2) (1:1): 0.5 mg/ml | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.8847 mL | 9.4233 mL | 18.8466 mL |
| 5 mM | 0.3769 mL | 1.8847 mL | 3.7693 mL |
| 10 mM | 0.1885 mL | 0.9423 mL | 1.8847 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 18 reference(s) in Google Scholar.)
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