Amine Coupling Kit

Cat. No.: A1507130

GRADE & PURITY BioReagent

Overview

  Amine Coupling Kit is a specialized set of reagents and materials used to covalent immobilization biomolecules (like antibodies, proteins, or peptides) carrying a primary amine group onto a carboxylated sensor surface for analysis using Surface Plasmon Resonance (SPR) biosensors.

The kit contains reagents sufficient for 30 to 50 immobilizations.

  The kit can be used with All Biacore™ systems except Biacore 4000, Biacore A100 and Biacore S51.

Contents and storage

A1507130	Component	1 kit	Storage
A1507130A	N-Hydroxysuccinimide (NHS)	115mg	2–8°C
A1507130B	1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)	750mg	2–8°C
A1507130C	1 M Ethanolamine hydrochloride-NaOH pH 8.5	10.5mL	2–8°C

Instructions for Use

1.     Reagent preparation

a)       Dissolve the EDC and NHS by adding 10.0 mL of filtered, deionized water to each vial.

b)       Cap vials tightly and agitate until the solids are completely dissolved.

c)       Dispense the EDC and NHS solutions separately in aliquots for storage.

d)       Store the aliquots of EDC and NHS frozen at -18°C or below. Ethanolamine may be stored at 2°C to 8°C.

Note: Use aliquots within two months.

2.     Handling

a)       Thaw frozen aliquots and agitate them gently to make sure that the solutions are homogeneous.

b)       For Biacore systems with autosampler:

Place the aliquots separately in the autosampler as instructed in the software.

c)       For Biacore systems without autosampler:

Mix equal volumes of EDC and NHS immediately before use.

3.     Suggested immobilization procedure

This guideline provides recommendations for immobilization of ligands by amine coupling to Biacore™ sensor chips. The user needs to optimize the reaction conditions based on the actual situation.

a)       Activate the surface by injecting a mixture of EDC/NHS (1:1) for 6–10 min (30 s for Sensor Chip PEG).

b)       Immobilize the ligand by injecting the ligand solution for 5–10 min.

c)       Deactivate excess reactive groups by injecting ethanolamine for 6–7 min.

Important considerations

1.     EDC and NHS Reagent solutions should be freshly prepared and mixed shortly before use. The efficiency of the immobilization will be reduced if solutions are not fresh.

2.     The EDC/NHS ester has a short half-life, so ligand should be injected shortly after activation

Storage and Shipping

Storage

Store at 2-8°C

Shipped In

Wet ice

Stability And Storage

Store at 2-8℃ long term (12 months).