| Product code | K AS-50I |
|---|---|
| Intended purpose | This kit contains all the necessary reagents to perform amine coupling via EDC/sulfo-NHS chemistry. It is mainly intended for use with 2D carboxyl-functionalized sensor chips, such as CMDP, HCP, CM-TEG and charge-reduced polycarboxylate HLC chips. Additionally, it is fully compatible with polycarboxylate hydrogel sensor chips. For immobilization using the Amine coupling kit AS, the ligand must contain primary amino groups. |
| Kit contents | Contains buffers and chemicals for 40–60 activations
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This kit provides the essential reagents for amine coupling via EDC/sulfo-NHS chemistry. It is specifically designed for use with XanTec SPR sensor chips carrying a (poly)carboxylate sensor matrix, including CMD, CMDP, HC, and HCP chips, as well as charge-reduced HLC sensor chips. Additionally, it is compatible with carboxy-functionalized sensor chips from other manufacturers. The kit includes sufficient material for approximately 50 injections. To achieve successful ligand immobilization using the Amine Coupling Kit AS, the ligand must contain primary amino groups. While primary amino groups are almost always present in proteins, small molecules, carbohydrates, and oligonucleotides may require additional functionalization to introduce these groups.
In contrast to the classical reagent NHS, sensor chip activation with sulfo-NHS preserves the negative charge of the activated carboxy groups. As a result, electrostatic preconcentration on the activated chip surface is less compromised, particularly when high EDC concentrations are used. However, it is important to note that the immobilization efficiency with sulfo-NHS may be lower than that when using NHS.
The EDC·HCl provided in this kit has undergone specialized in-house purification and testing to ensure superior performance in SPR applications. This is particularly important because even high-purity EDC·HCl often contains impurities, such as diamines, that can neutralize negative surface charges and deactivate sulfo-NHS esters on an activated chip surface. These impurities can significantly decrease immobilization efficiency, especially at high EDC concentrations; in extreme cases, they may cause complete deactivation of the chip surface.
For optimal results, EDC should be used within the concentration ranges specified in Table 1. It is worth noting that the ideal concentrations vary significantly depending on the sensor chip matrix.
Table 1: Recommended EDC concentration ranges for XanTec (poly)carboxylate sensor chips, using sulfo-NHS.
| Sensor chip type | Min. EDC | Max. EDC |
|---|---|---|
| HCP | 0.5 % | 2.0 % |
| HC | 0.5 % | 5.0 % |
| CMDP | 1.0 % | 5.0 % |
| CMD | 0.5 % | 5.0 % |
| HLC | 0.5 % | 5.0 % |
Aqueous EDC solutions are susceptible to hydrolysis. Therefore, Activation Buffer should be frozen in small aliquots, and the corresponding amount of solid EDC·HCl should be dissolved immediately in the buffer before use. Because EDC is sensitive to humidity, the reagent bottle should be stored at -20 °C or lower in a desiccated container, brought to room temperature before opening, and opened for as short a time as possible, ideally under an inert gas atmosphere.
Alternatively, solid EDC·HCl can be dissolved in ultrapure water to prepare a 10 % EDC solution (521 mM), which can then be divided into aliquots for storage at -20 °C or lower. These aliquots remain stable for up to two months when kept frozen. Immediately before use, thaw a frozen aliquot to room temperature, gently shake it, and combine it with thawed Activation Buffer. This mixture can then be used for activating the sensor chip.
For detailed activation protocols, please refer to the specific instructions provided for the respective sensor chip types.
Instructions for Use – 2D Carboxylate Sensor Chips
Instructions for Use – HLC Sensor Chips
Instructions for Use – Polycarboxylate Hydrogel Sensor Chips
V. 10/24a
For in-vitro use only. Not for use in clinical diagnostic procedures.