School of Pharmaceutical Sciences, Tsinghua University, 100084 Beijing, China.
Tsinghua-Peking Center for Life Sciences, Tsinghua University, 100084 Beijing, China.
STAR Protoc. 2021 Dec 23;3(1):101061. doi: 10.1016/j.xpro.2021.101061. eCollection 2022 Mar 18.
Toll-like receptor 8 (TLR8) is a pattern recognition receptor that senses RNA degradation products and initiates immune responses. TLR8 overactivation is associated with autoimmune diseases. Herein, we describe the evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase (SEAP) assay, WST assay, ITC and immunoblotting. These assays can facilitate the development of TLR8 antagonists; this protocol can also be adapted to analyze agonists and antagonists for other TLRs. For complete details on the use and execution of this protocol, please refer to Hu et al. (2018).
Toll 样受体 8（TLR8）是一种模式识别受体，可识别 RNA 降解产物并引发免疫反应。TLR8 的过度激活与自身免疫性疾病有关。在此，我们通过 SEAP 测定、WST 测定、ITC 和免疫印迹描述了 TLR8 拮抗剂在 HEK-Blue 细胞中的评估和验证。这些测定方法可以促进 TLR8 拮抗剂的开发；该方案也可以适应分析其他 TLR 的激动剂和拮抗剂。有关此方案使用和执行的完整详细信息，请参阅 Hu 等人。（2018 年）。
