logo
Products
Services
Resources
Selection Guides
About
Labeling Azide-Modified Biomolecules with Fluorescent Dye Alkynes
Labeling Oligonucleotides with Dye Alkynes
  1. Prepare the following stock solutions:
    • 200 mM THPTA [tris(3- hydroxypropyltriazolylmethyl)amine)] in water
    • 100 mM CuSO4 in water
    • Azide-modified oligo in water (as concentrated as possible, e.g., >10 mg/ml)
    • 100 mM sodium ascorbate in water
    • 10 mM dye alkyne in DMSO or water (see our website for recommended solvent)
  2. Mix and vortex well CuSO4 with THPTA in a 1:2 ratio for several minutes before the reaction. This working solution is stable for several weeks when frozen.
  3. To the azide-modified oligo solution, add an excess of dye alkyne (2-5 equivalents by molar ratio).
  4. Add 5 equivalents of THPTA/CuSO4working solution (from Step 1)
  5. Add 10-30 equivalents of sodium
  6. Stir, vortex or shake the reaction mixture at room temperature for 30-60 minutes.
  7. Ethanol-precipitate the oligo or purify it by your desired method (e.g., HPLC).
Labeling Peptides with Dye Alkynes
  1. Prepare the following stock solutions:
    • 200 mM THPTA ligand in water
    • 100 mM CuSO4 in water
    • Azide-modified peptide in water or DMF (depending on your peptide solubility, >10 mg/ml if possible)
    • 100 mM sodium ascorbate in water
    • 10 mM dye alkyne in DMSO or water (see our website for recommended solvent)
  2. Incubate CuSO4 with THPTA ligand in a 1:2 ratio several minutes before the reaction. This solution is stable for several weeks when frozen.
  3. To the azide-modified peptide solution, add an excess of dye alkyne (5-10 equivalents by molar ratio).
  4. Add 5-10 equivalents of THPTA/CuSO4.
  5. Add 10-20 equivalents of sodium.
  6. Stir, vortex or shake the reaction mixture at room temperature for 30-60 minutes.
  7. Purify your desired peptide by HPLC.
Labeling Small Organic Alkyne Molecules with Dye Alkynes
  1. Prepare the following stock solutions:
    • 200 mM THPTA ligand in water
    • 100 mM CuSO4 in water
    • Alkyne compound in water or DMF (depending on your compound solubility, >10 mg/ml if possible,)
    • 100 mM sodium ascorbate in water
    • 10 mM dye alkyne in DMSO or water (see our website for recommended solvent).
  2. Incubate CuSO4 with THPTA ligand in a 1:2 ratio several minutes before the reaction. This solution is stable for several weeks when frozen.
  3. To the alkyne solution, add an excess of dye alkyne (5-10 equivalents by molar ratio).
  4. Add 25 equivalents of THPTA/CuSO4.
  5. Add 50 equivalents of sodium
  6. Stir the reaction mixture at room temperature for 30-60 minutes.
  7. Purify your desired molecule by chromatography or other methods.
Labeling Biopolymers with Dye Alkynes
  1. Prepare the following stock solutions:
    • 200 mM THPTA ligand in water
    • 100 mM CuSO4 in water
    • Azide-modified biopolymer in water (as concentrated as possible, e.g., >5 mg/ml)
    • 100 mM sodium ascorbate in water
    • 10 mM dye alkyne in DMSO or water (see our website for recommended solvent).
  2. Incubate CuSO4 with THPTA ligand in a 1:2 ratio several minutes before the reaction. This solution is stable for several weeks when frozen.
  3. To the azide-modified biopolymer solution, add an excess of dye alkyne (Loading ratio: 5-20 dye alkyne/alkyne).
  4. Add 5 molar equivalents (referenced to dye alkyne) of THPTA/CuSO4.
  5. Add 10 equivalents of sodium ascorbate (referenced to dye alkyne).
  6. Stir, vortex or shake the reaction mixture at room temperature for 30-60 minutes.
  7. Purify your desired molecule by gel filtration or dialysis.
This document (02.0117.180413r1) was last updated on Mon Oct 13 2025. All trademarks and registered trademarks mentioned herein are the property of their respective owners.
Labeling Azide-Modified Biomolecules with Fluorescent Dye Alkynes
Labeling Oligonucleotides with Dye Alkynes
Labeling Peptides with Dye Alkynes
Labeling Small Organic Alkyne Molecules with Dye Alkynes
Labeling Biopolymers with Dye Alkynes