NTA Chips for SPR: Difference between revisions
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Nieba et al. <cite>#his1</cite> | 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 <cite>#his1</cite>. | ||
Affinity of singly his-tagged proteins to the NiNTA complex is insufficient to immobilize such proteins. | |||
Also, Nieba et al.<cite>#his1</cite> studied several parameters affecting level of ligand immobilization: | |||
*conditions for NTA surface activation with Ni<sup>2+</sup>: [NiCl<sub>2</sub>], [NaCl] and pH of activation buffer | *conditions for NTA surface activation with Ni<sup>2+</sup>: [NiCl<sub>2</sub>], [NaCl] and pH of activation buffer | ||
*effects of pH, [NaCl] and detergent (tween) concentration in the ligand immobilization buffer | *effects of pH, [NaCl] and detergent (tween) concentration in the ligand immobilization buffer | ||
Revision as of 19:39, 21 March 2008
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 [1].
Affinity of singly his-tagged proteins to the NiNTA complex is insufficient to immobilize such proteins.
Also, Nieba et al.[1] studied several 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 and number of his-tags per ligand molecule
Also surface regeneration conditions were optimized.
