Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine

This protocol describes how to characterize α-Smooth muscle actin (αSMA) spatiotemporal expression during mouse small intestinal development. Specific tissue fixation preserves αSMA arrangement in low αSMA expressing cells that are conventionally undetectable under αSMA immunofluorescent stain due t...

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Detalles Bibliográficos
Autores principales: Hu, Shing, Sevier, Carolyn S., Kurpios, Natasza A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9284457/
https://www.ncbi.nlm.nih.gov/pubmed/35810413
http://dx.doi.org/10.1016/j.xpro.2022.101524
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author Hu, Shing
Sevier, Carolyn S.
Kurpios, Natasza A.
author_facet Hu, Shing
Sevier, Carolyn S.
Kurpios, Natasza A.
author_sort Hu, Shing
collection PubMed
description This protocol describes how to characterize α-Smooth muscle actin (αSMA) spatiotemporal expression during mouse small intestinal development. Specific tissue fixation preserves αSMA arrangement in low αSMA expressing cells that are conventionally undetectable under αSMA immunofluorescent stain due to inappropriate fixative-caused artificial actin depolymerization. Parallel analysis of αSMA carbonylation allows estimation of oxidative damage in gut muscular lineage. This approach improves the molecular specificity offered by commercialized kits that estimate total protein carbonyl level in cell lysates without protein specificity. For complete details on the use and execution of this protocol, please refer to Hu et al. (2021).
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spelling pubmed-92844572022-07-16 Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine Hu, Shing Sevier, Carolyn S. Kurpios, Natasza A. STAR Protoc Protocol This protocol describes how to characterize α-Smooth muscle actin (αSMA) spatiotemporal expression during mouse small intestinal development. Specific tissue fixation preserves αSMA arrangement in low αSMA expressing cells that are conventionally undetectable under αSMA immunofluorescent stain due to inappropriate fixative-caused artificial actin depolymerization. Parallel analysis of αSMA carbonylation allows estimation of oxidative damage in gut muscular lineage. This approach improves the molecular specificity offered by commercialized kits that estimate total protein carbonyl level in cell lysates without protein specificity. For complete details on the use and execution of this protocol, please refer to Hu et al. (2021). Elsevier 2022-07-09 /pmc/articles/PMC9284457/ /pubmed/35810413 http://dx.doi.org/10.1016/j.xpro.2022.101524 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Hu, Shing
Sevier, Carolyn S.
Kurpios, Natasza A.
Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
title Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
title_full Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
title_fullStr Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
title_full_unstemmed Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
title_short Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
title_sort protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9284457/
https://www.ncbi.nlm.nih.gov/pubmed/35810413
http://dx.doi.org/10.1016/j.xpro.2022.101524
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