FAM azide, 6-isomer

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Prepare Working Solutions
1.1 FAM azide, 6-isomer stock solution: Prepare a 10 mM stock solution with anhydrous DMSO. Aliquot the solution and store it at -20°C in the dark; avoid repeated freeze-thaw cycles.
1.2 Other essential reagents:
Copper complex: For example, mix a 100 mM aqueous CuSO₄ solution with a 200 mM aqueous THPTA ligand solution at a volume ratio of 1:2, then vortex to mix thoroughly for later use. As a stabilizing ligand for copper ions, THPTA protects biomolecules and accelerates the reaction.
Reducing agent: 100 mM aqueous sodium ascorbate solution, which requires fresh preparation.
Sample to be labeled: A solution of target molecules with terminal alkynes (such as oligonucleotides, peptides, proteins, etc.).

2. Perform Click Chemistry Reaction
2. Mix the following components in a microcentrifuge tube:
Alkynylated sample to be labeled (e.g., concentration >10 mg/mL);
10 mM FAM azide, 6-isomer stock solution (2-10 molar equivalents in excess relative to alkynes);
Add 5-10 equivalents (relative to dye azide) of the pre-mixed CuSO₄/THPTA solution;
Add 10-30 equivalents (relative to dye azide) of freshly prepared 100 mM sodium ascorbate solution to initiate the reaction;
Vortex briefly to mix, then incubate the reaction at room temperature in the dark for 30-60 minutes. For some samples, extended reaction time or temperature optimization may be required.

3. Post-Reaction Purification
After the reaction completes, separate the labeled product from excess free dye using an appropriate method.
For oligonucleotides and peptides: Purification is achievable via ethanol precipitation or HPLC.
For biomacromolecules such as proteins: Purification is achievable via gel filtration chromatography or dialysis.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.