Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals

Previous immunostaining protocols are highly specific for model organisms and often not suitable for diverse specimens that are non-perfused and over-fixed (i.e., tissues sitting in fixatives for months/year). Here, we present an immunofluorescence protocol for localizing protein targets in brain ti...

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Detalles Bibliográficos
Autores principales: Ciani, Caterina, Pistorio, Giulio, Mearelli, Marika, Falcone, Carmen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10432796/
https://www.ncbi.nlm.nih.gov/pubmed/37561635
http://dx.doi.org/10.1016/j.xpro.2023.102482
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author Ciani, Caterina
Pistorio, Giulio
Mearelli, Marika
Falcone, Carmen
author_facet Ciani, Caterina
Pistorio, Giulio
Mearelli, Marika
Falcone, Carmen
author_sort Ciani, Caterina
collection PubMed
description Previous immunostaining protocols are highly specific for model organisms and often not suitable for diverse specimens that are non-perfused and over-fixed (i.e., tissues sitting in fixatives for months/year). Here, we present an immunofluorescence protocol for localizing protein targets in brain tissue from 11 model and non-model mammals. We describe preparation of both fresh and fixed tissues including steps for deparaffinization, fixation, and cryoprotection. We then detail immunofluorescence procedures including antigen retrieval, reducing autofluorescence, nuclear staining, mounting, and image collection.
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spelling pubmed-104327962023-08-18 Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals Ciani, Caterina Pistorio, Giulio Mearelli, Marika Falcone, Carmen STAR Protoc Protocol Previous immunostaining protocols are highly specific for model organisms and often not suitable for diverse specimens that are non-perfused and over-fixed (i.e., tissues sitting in fixatives for months/year). Here, we present an immunofluorescence protocol for localizing protein targets in brain tissue from 11 model and non-model mammals. We describe preparation of both fresh and fixed tissues including steps for deparaffinization, fixation, and cryoprotection. We then detail immunofluorescence procedures including antigen retrieval, reducing autofluorescence, nuclear staining, mounting, and image collection. Elsevier 2023-08-09 /pmc/articles/PMC10432796/ /pubmed/37561635 http://dx.doi.org/10.1016/j.xpro.2023.102482 Text en © 2023 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
Ciani, Caterina
Pistorio, Giulio
Mearelli, Marika
Falcone, Carmen
Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
title Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
title_full Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
title_fullStr Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
title_full_unstemmed Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
title_short Immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
title_sort immunofluorescence protocol for localizing protein targets in brain tissue from diverse model and non-model mammals
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10432796/
https://www.ncbi.nlm.nih.gov/pubmed/37561635
http://dx.doi.org/10.1016/j.xpro.2023.102482
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