NTA Chips for SPR: Difference between revisions
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==Introduction== | |||
NTA sensor chips can be successfully used for immobilization of molecules carrying more then two his-tags, as shown by Nieba et al. <cite>#his1</cite>, e.g. with oligomeric proteins. Sometimes immobilization can even succeed with two his-tags per molecule, while affinity of singly his-tagged proteins to the NiNTA complex is insufficient to immobilize such proteins. | NTA sensor chips can be successfully used for immobilization of molecules carrying more then two his-tags, as shown by Nieba et al. <cite>#his1</cite>, e.g. with oligomeric proteins. Sometimes immobilization can even succeed with two his-tags per molecule, while affinity of singly his-tagged proteins to the NiNTA complex is insufficient to immobilize such proteins. | ||
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*effect of Na<sub>2</sub>EDTA (0-300 mM) in the running and ligand buffers | *effect of Na<sub>2</sub>EDTA (0-300 mM) in the running and ligand buffers | ||
*concentration of his-tagged ligand | *concentration of his-tagged ligand | ||
==Chip activation== | |||
Surface regeneration conditions were optimized as well. | Surface regeneration conditions were optimized as well. |
Revision as of 15:39, 24 March 2008
Introduction
NTA sensor chips can be successfully used for immobilization of molecules carrying more then two his-tags, as shown by Nieba et al. [1], e.g. with oligomeric proteins. Sometimes immobilization can even succeed with two his-tags per molecule, while affinity of singly his-tagged proteins to the NiNTA complex is insufficient to immobilize such proteins.
Also, Nieba et al.[1] studied several other parameters affecting level of ligand immobilization:
- conditions for NTA surface activation with Ni2+: [NiCl2], [NaCl] and pH of activation buffer
- effects of pH, [NaCl] and detergent (tween) concentration in the ligand immobilization buffer
- effect of Na2EDTA (0-300 mM) in the running and ligand buffers
- concentration of his-tagged ligand
Chip activation
Surface regeneration conditions were optimized as well.