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Instructions
for use

Amine coupling kit AZ, zwitterionic surfaces (PDF download)

Product description

Product code K AZ-30I
Intended purpose This kit includes all essential reagents required for amine coupling via EDC/NHS chemistry on zwitterionic surfaces. It is intended for use with ZC sensor chips from XanTec, which are coated with a zwitterionic polymer containing an almost equimolar ratio of carboxylic acids and tertiary amines. A prerequisite for use of this kit is the presence of at least one primary amino group on the ligand.
Kit contents Contains buffers and chemicals for about 30 activations
  • Solid EDC·HCl, purity >99 % SPR-grade, 1.5 g
  • Activation buffer, 0.05 M MES, 0.1 M NHS, pH 5.0, 50 mL
  • 10× 0.01 M MES immobilisation buffer, pH 6.5, 50 mL
  • Borate elution buffer: 0.1 M sodium borate, 1 M NaCl, pH 9.0, 50 mL
  • Quenching buffer A, 1 M glycine·HCl, 0.01 M MES, 0.01 % thiomersal, pH 6.5, 50 mL
  • Quenching buffer C, 1 M N,N-dimethylethylenediamine (DMEN·HCl), pH 6.5, 50 mL
Storage
  • Solid EDC·HCl: Store at -20 °C, desiccated over molecular sieve 4A
  • Activation buffer: Store at -20 °C
  • 10× MES immobilisation buffer: Store at -20 °C
  • Borate elution buffer: Store at room temperature or -20 °C
  • Quenching buffers A and B: Store at room temperature or -20 °C
Related products
  • ZC sensor chip (e.g., ZC80M, ZC30M)
  • ZCC sensor chip (e.g., ZCC150D)

Instructions

This kit provides the essential reagents for amine coupling via EDC/NHS chemistry in combination with ZC sensor chips. The amount of EDC·HCl and the immobilization and quenching buffers have been modified to account for the particular requirements associated with activation of ZC sensor chips. To achieve successful ligand immobilization using the Amine Coupling Kit AZ, the ligand must contain at least one primary amino group.

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 NHS esters on the 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.

EDC preparation

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 ultra-pure water to prepare a 1 M EDC solution, which can then be divided into separate 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 the defrosted Activation buffer. This mixture can then be used for activating the ZC sensor chip.

Related documents

Instructions for Use – ZC Sensor Chips

V. 10/24a

For in-vitro use only. Not for use in clinical diagnostic procedures.