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Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies
Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissue...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9852956/ https://www.ncbi.nlm.nih.gov/pubmed/36853733 http://dx.doi.org/10.1016/j.xpro.2022.102033 |
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author | de Faria, Isaque J.S. Imler, Jean-Luc Marques, João T. |
author_facet | de Faria, Isaque J.S. Imler, Jean-Luc Marques, João T. |
author_sort | de Faria, Isaque J.S. |
collection | PubMed |
description | Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).(1) |
format | Online Article Text |
id | pubmed-9852956 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-98529562023-01-21 Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies de Faria, Isaque J.S. Imler, Jean-Luc Marques, João T. STAR Protoc Protocol Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).(1) Elsevier 2023-01-14 /pmc/articles/PMC9852956/ /pubmed/36853733 http://dx.doi.org/10.1016/j.xpro.2022.102033 Text en © 2022 The Authors 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 de Faria, Isaque J.S. Imler, Jean-Luc Marques, João T. Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies |
title | Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies |
title_full | Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies |
title_fullStr | Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies |
title_full_unstemmed | Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies |
title_short | Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies |
title_sort | protocol for the analysis of double-stranded rnas in virus-infected insect cells using anti-dsrna antibodies |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9852956/ https://www.ncbi.nlm.nih.gov/pubmed/36853733 http://dx.doi.org/10.1016/j.xpro.2022.102033 |
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