Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: de Faria, Isaque J.S., Imler, Jean-Luc, Marques, João T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
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
_version_ 1784872783790997504
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
work_keys_str_mv AT defariaisaquejs protocolfortheanalysisofdoublestrandedrnasinvirusinfectedinsectcellsusingantidsrnaantibodies
AT imlerjeanluc protocolfortheanalysisofdoublestrandedrnasinvirusinfectedinsectcellsusingantidsrnaantibodies
AT marquesjoaot protocolfortheanalysisofdoublestrandedrnasinvirusinfectedinsectcellsusingantidsrnaantibodies