| Assay Method Information | |
| | AlphaLISA |
| Description: | The ability of compounds to inhibit the binding of IL-17A to IL-17RA may be assessed with an AlphaLISA assay. Expression and purification of protein reagents IL17A and IL17RA for alphaLISA may be carried out essentially as follows. A construct of cytokine IL-17A (Reference sequence NP_002181) residues 1-155 with a C-terminal AVI-tag, followed by a His tag, is expressed in insect cells, together with BirA for in vivo biotinylation. Receptor IL-17RA (NP_055154) residues 1-317 with mutations N206D/N265D is expressed in insect cells with a C-terminal thrombin cleavage site followed by a FLAG-tag. IL-17A and IL-17RA are each PCR amplified and TOPO cloned into custom TOPO adapted pFastBac vectors (Thermo Fisher Scientific. Carlsbad, Calif.). |
| Affinity data for this assay | |
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