DBCO-Sulfo-NHS Ester

Catalog # Pkg Size Price(USD) Quantity
A124-10 10 mg $95.00
A124-25 25 mg $195.00
A124-100 100 mg $495.00
A124-500 500 mg $1,695.00

DBCO-Sulfo-NHS Ester is water-soluble reagent that enables simple and efficient labeling of antibodies, proteins and any other primary amine-containing macromolecules with DBCO moiety in 100% aqueous buffers.
DBCO-Sulfo-NHS Ester is a reagent of choice for applications that cannot tolerate organic co-solvents or are complicated by their inclusion. Specific labeling of cell surface proteins is another common application for these uniquely water-soluble and membrane impermeable reagents.

 

Protein Labeling Calculator

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** Please fill all the required input **

  • Volume of DMSO required to make a stock solution (µL)

  • Volume of protein to be labeled (µL)

  • Volume of biotin stock solution to add (µL)

  • Percent DMSO in Reaction Mixture (%)

Caution - Volume of DSMO solvent added to protein must always be <5% total volume of reaction

Important Product Information

  • DBCO-sulfo-NHS ester is moisture-sensitive. To avoid moisture condensation onto the product always let vial come to room temperature before opening; be careful to limit exposure to moisture and restore under an inert atmosphere. The NHS-ester moiety readily hydrolyzes and becomes non-reactive; therefore, prepare stock solutions immediately before use. Stock solutions in anhydrous solvents can be kept for several days (freeze when not in use).
  • Hydrolysis of DBCO-sulfo-NHS ester is a competing reaction. Conjugation with primary amines of proteins/peptides (i.e., acylation) is favored at near neutral pH (6-9) and with concentrated protein solutions. For conjugation, use non-amine-containing buffers at pH 7-9 such as PBS (20 mM sodium phosphate, 150 mM sodium chloride, pH 7.4); 20 mM HEPES; 100 mM carbonate/biocarbonate; or 50 mM borate buffer.
  • Do not use buffers that contain primary amines, (e.g., Tris, glycine).
  • Avoid buffers that contain azides, which can react with DBCO.
  • Dissolve DBCO-Sulfo-NHS ester in a aqueous buffer or dry water-miscible organic solvent such as DMSO or DMF before diluting in final reaction buffer.
  • Reactions with DBCO and azides are more efficient at high concentrations and temperatures (i.e., 2-37°C). Typical reaction times are less than 4 hours; however, incubating for longer can improve efficiency.

 

Procedure for Sample Labeling

Additional Materials Required

  • Aqueous buffer or water-miscible organic solvent such as dimethyl sulfoxide (DMSO) or dimethyl formamide (DMF)
  • Reaction buffer: Phosphate-buffered saline (PBS) or other buffer at pH 5-9
  • Quenching buffer: 1 M Tris∙HCl, Ph 8.0
  • Spin Desalting Columns

Protein Derivitization

  • Prepare proteins in PBS.
  • Immediately before use, prepare 10 mM of the DBCO-sulfo-NHS ester in aqueous buffer, DMSO or DMF.
  • Add the NHS reagent to the protein sample at a final concentration of 0.5-2 mM. If the protein concentration is ≥ 5 mg/ml, use a 10-fold molar excess of the reagent. For samples < 5 mg/ml, use a 20- to 50-fold molar excess.
  • Incubate the reaction at room temperature for 30 minutes or on ice for 2 hours.
  • Stop the reaction by adding Quenching Buffer to a final concentration of 50-100 mM Tris.
  • Incubate the reaction at room temperature for 5 minutes or on ice for 15 minutes.
  • Remove non-reactive reagent by dialysis or desalting.

 

Copper-free Click Reaction

  • Prepare the azide-containing sample in reaction buffer.
  • Add DBCO-protein conjugate to azide-containing sample.

Recommendation: Add 1 mole equivalent of limiting reagent to 1.5-3.0 mol equivalents of highest abundance reagent.

  • Incubate the reaction at room temperature for 4-12 hour. Incubation at 40C requires 2-12 hours.
  • The reaction is now ready for purification.

Download pdf

1. Wang, J.S., et al. (2015). High-Density PEO-b-DNA Brushes on Polymer Particles for Colloidal Superstructures. Chem. Mater., 27(24), 8337-8344.

2. Guhuan, L., et al. (2015). Hyperbranched Self-Immolative Polymers (hSIPs) for Programmed Payload Delivery and Ultrasensitive Detection. J. Am. Chem. Soc., 137(36), 11645-11655.

3. Sukwon, J., et al. (2013). Facile Strategy for Protein Conjugation with Chitosan-Poly(ethylene glycol) Hybrid Microparticle Platforms via Strain-Promoted Alkyne-Azide Cycloaddition (SPAAC) Reaction. Biomacromolecules, 14(11), 3892-3902.

4. Sukwon, J., et al. (2012). Fabrication of Chitosan-Poly(ethylene glycol) Hybrid Hydrogel Microparticles via Replica Molding and Its Application toward Facile Conjugation of Biomolecules. Langmuir, 28(49), 17061-17070.