| Assay Method Information | |
| | Surface Plasmon Resonance (SPR) Assay |
| Description: | Pure His-cyclophilins were immobilized and covalently stabilized on the NTA sensor chip according to the protocol described in Thermo-kinetic analysis space expansion for cyclophilin-ligand interactions—identification of a new nonpeptide inhibitor using Biacore T200. Using 200 nM concentrations of each protein, in Running Buffer (PBS, pH 7.4; 0.05% surfactant P20, 2% v/v ethanol; 50 μM EDTA), at 30 μl min−1 with 60 second contact times on the activated NTA surfaces. This gave signals of 1,921 RU for His-CypA, 1932 RU for His-CypB and 1,397 RU for His-CypD. Specific surface protein activity was assayed by passing saturating amounts of CsA (2 μM) in Running Buffer over these surfaces; values of 94.1%, 95.5% and 95.6% activity were obtained for His-CypA, -B and -D, respectively. |
| Affinity data for this assay | |
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