Mass isolation of staged Drosophila pupal intestines for analysis of protein ubiquitylation.
Cell Biology
Developmental biology
Microscopy
Model Organisms
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
10 Nov 2023
10 Nov 2023
Historique:
received:
30
08
2023
revised:
04
10
2023
accepted:
26
10
2023
medline:
11
11
2023
pubmed:
11
11
2023
entrez:
11
11
2023
Statut:
aheadofprint
Résumé
Large quantities of developmentally synchronized pupal intestines are required for biochemistry experiments. Here, we present a protocol for the mass isolation of staged pupal intestines during Drosophila melanogaster development based on buoyancy in sucrose for biochemical evaluation of protein ubiquitylation. We describe steps for crossing flies, preparation of samples, immunoprecipitation of proteins from staged isolated tissues, and analysis of samples by western blot. This protocol can be extended to investigate biochemical changes in other tissues. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023).
Identifiants
pubmed: 37950865
pii: S2666-1667(23)00680-9
doi: 10.1016/j.xpro.2023.102713
pmc: PMC10682243
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
102713Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM131689
Pays : United States
Informations de copyright
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.
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