Human IL17 kit
No-wash kit to quantify released Human IL17
The Human IL17A/IL17RA binding kit is designed to identify human IL17A/IL17RA inhibitors.
Interleukin 17A , a pro-inflammatory cytokine, belongs to a family of six cytokines; IL-17A, IL-17B, IL-17C, IL-17D, IL-E, and IL-17F. IL-17A binds a heterodimeric cell surface receptor (IL-17RA/IL-17RC), triggering a signaling cascade that results in NF-κB activation which leads to the stimulation of the pro-inflammatory cytokines IL-6 and IL-8. Although this cytokine exerts a protective role against many pathogens, the overproduction of IL-17A is associated with autoimmune diseases such as psoriasis, rheumatid arthritis, and Crohn's Disease.
The Human IL17RA/IL17RA binding kit can be run in 96- or 384-well low volume white plates (20 µL final). As described here, samples or standards are dispensed directly into the assay plate. The Tag1-IL17A protein and Tag2-IL17RA proteins are then added, followed by the dispensing of the HTRF reagents: the anti Tag1 reagent labeled with europium cryptate and the anti Tag2 antibody labeled with XL665. The reagents labeled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
The inhibitory effects of unlabeled IL17A and IL17R, and human anti-IL17A antibody, were tested. No inhibition was observed using an anti EGFR antibody (cetuximab), confirming the specificity of the kit for the study of IL17A/IL17RA interaction.
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