Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay

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 embryoni...

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Autores principales: Hu, Zhenyi, Zhang, Tuan, Jiang, Shuangshuang, Yin, Hang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715332/
https://www.ncbi.nlm.nih.gov/pubmed/35005643
http://dx.doi.org/10.1016/j.xpro.2021.101061
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author Hu, Zhenyi
Zhang, Tuan
Jiang, Shuangshuang
Yin, Hang
author_facet Hu, Zhenyi
Zhang, Tuan
Jiang, Shuangshuang
Yin, Hang
author_sort Hu, Zhenyi
collection PubMed
description 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).
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spelling pubmed-87153322022-01-06 Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay Hu, Zhenyi Zhang, Tuan Jiang, Shuangshuang Yin, Hang STAR Protoc Protocol 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). Elsevier 2021-12-23 /pmc/articles/PMC8715332/ /pubmed/35005643 http://dx.doi.org/10.1016/j.xpro.2021.101061 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Hu, Zhenyi
Zhang, Tuan
Jiang, Shuangshuang
Yin, Hang
Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay
title Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay
title_full Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay
title_fullStr Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay
title_full_unstemmed Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay
title_short Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay
title_sort protocol for evaluation and validation of tlr8 antagonists in hek-blue cells via secreted embryonic alkaline phosphatase assay
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715332/
https://www.ncbi.nlm.nih.gov/pubmed/35005643
http://dx.doi.org/10.1016/j.xpro.2021.101061
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