| Assay Method Information | |
| | Biolayer Interferometry (BLI) Assay |
| Description: | The experiments were conducted using the Octet Red System (Forte bio, MenloPark, CA). Biotinylated peptides were immobilized on kinetics grade Streptavidin biosensors. The peptides, coupled to the biosensors, were titrated with σB at increasing concentrations in a buffer with 25 mM phosphate and 125 mM NaCl (PBS) with 0.1 mg/mL BSA and 0.05% (v/v) Tween 20; 0.01% BSA and 0.05% (v/v) Tween 20 were used to prevent nonspecific interaction of σB with the sensor surface. The analyte was subsequently allowed to dissociate in PBS with 0.1 mg/mL BSA and 0.05% (v/v) Tween 20. The ligands were regenerated for reuse by washing the bound analyte (σB) with 1 M MgCl2. A loaded biosensor with only interaction buffer was used as a control for nonspecific interactions. |
| Affinity data for this assay | |
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