Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation

Isolation of skeletal muscles allows for the exploration of many complex diseases. Here, we present a protocol for isolating mice skeletal muscle myoblasts and myotubes that have been differentiated through antibody validation. We describe steps for collecting and preparing murine skeletal tissue, m...

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Autores principales: Stephens, Dominique C., Mungai, Margaret, Crabtree, Amber, Beasley, Heather K., Garza-Lopez, Edgar, Vang, Larry, Neikirk, Kit, Vue, Zer, Vue, Neng, Marshall, Andrea G., Turner, Kyrin, Shao, Jian-qiang, Sarker, Bishnu, Murray, Sandra, Gaddy, Jennifer A., Davis, Jamaine, Damo, Steven M., Hinton, Antentor O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10663959/
https://www.ncbi.nlm.nih.gov/pubmed/37938976
http://dx.doi.org/10.1016/j.xpro.2023.102591
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author Stephens, Dominique C.
Mungai, Margaret
Crabtree, Amber
Beasley, Heather K.
Garza-Lopez, Edgar
Vang, Larry
Neikirk, Kit
Vue, Zer
Vue, Neng
Marshall, Andrea G.
Turner, Kyrin
Shao, Jian-qiang
Sarker, Bishnu
Murray, Sandra
Gaddy, Jennifer A.
Davis, Jamaine
Damo, Steven M.
Hinton, Antentor O.
author_facet Stephens, Dominique C.
Mungai, Margaret
Crabtree, Amber
Beasley, Heather K.
Garza-Lopez, Edgar
Vang, Larry
Neikirk, Kit
Vue, Zer
Vue, Neng
Marshall, Andrea G.
Turner, Kyrin
Shao, Jian-qiang
Sarker, Bishnu
Murray, Sandra
Gaddy, Jennifer A.
Davis, Jamaine
Damo, Steven M.
Hinton, Antentor O.
author_sort Stephens, Dominique C.
collection PubMed
description Isolation of skeletal muscles allows for the exploration of many complex diseases. Here, we present a protocol for isolating mice skeletal muscle myoblasts and myotubes that have been differentiated through antibody validation. We describe steps for collecting and preparing murine skeletal tissue, myoblast cell maintenance, plating, and cell differentiation. We then detail procedures for cell incubation, immunostaining, slide preparation and storage, and imaging for immunofluorescence validation.
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spelling pubmed-106639592023-11-07 Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation Stephens, Dominique C. Mungai, Margaret Crabtree, Amber Beasley, Heather K. Garza-Lopez, Edgar Vang, Larry Neikirk, Kit Vue, Zer Vue, Neng Marshall, Andrea G. Turner, Kyrin Shao, Jian-qiang Sarker, Bishnu Murray, Sandra Gaddy, Jennifer A. Davis, Jamaine Damo, Steven M. Hinton, Antentor O. STAR Protoc Protocol Isolation of skeletal muscles allows for the exploration of many complex diseases. Here, we present a protocol for isolating mice skeletal muscle myoblasts and myotubes that have been differentiated through antibody validation. We describe steps for collecting and preparing murine skeletal tissue, myoblast cell maintenance, plating, and cell differentiation. We then detail procedures for cell incubation, immunostaining, slide preparation and storage, and imaging for immunofluorescence validation. Elsevier 2023-11-07 /pmc/articles/PMC10663959/ /pubmed/37938976 http://dx.doi.org/10.1016/j.xpro.2023.102591 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
Stephens, Dominique C.
Mungai, Margaret
Crabtree, Amber
Beasley, Heather K.
Garza-Lopez, Edgar
Vang, Larry
Neikirk, Kit
Vue, Zer
Vue, Neng
Marshall, Andrea G.
Turner, Kyrin
Shao, Jian-qiang
Sarker, Bishnu
Murray, Sandra
Gaddy, Jennifer A.
Davis, Jamaine
Damo, Steven M.
Hinton, Antentor O.
Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
title Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
title_full Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
title_fullStr Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
title_full_unstemmed Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
title_short Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
title_sort protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10663959/
https://www.ncbi.nlm.nih.gov/pubmed/37938976
http://dx.doi.org/10.1016/j.xpro.2023.102591
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