Cargando…
An antibody toolkit for the study of membrane traffic in Drosophila melanogaster
The use of Drosophila melanogaster as a model organism has been pivotal to understanding the developmental processes of metazoans. However, the use of flies for studying subcellular organization is hampered by a paucity of reliable reagents to label specific organelles. Here, we describe the generat...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists Ltd
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4958275/ https://www.ncbi.nlm.nih.gov/pubmed/27256406 http://dx.doi.org/10.1242/bio.018937 |
_version_ | 1782444286170628096 |
---|---|
author | Riedel, Falko Gillingham, Alison K. Rosa-Ferreira, Cláudia Galindo, Antonio Munro, Sean |
author_facet | Riedel, Falko Gillingham, Alison K. Rosa-Ferreira, Cláudia Galindo, Antonio Munro, Sean |
author_sort | Riedel, Falko |
collection | PubMed |
description | The use of Drosophila melanogaster as a model organism has been pivotal to understanding the developmental processes of metazoans. However, the use of flies for studying subcellular organization is hampered by a paucity of reliable reagents to label specific organelles. Here, we describe the generation of mouse monoclonal antibodies against a set of markers of the secretory and endocytic pathways, along with goat polyclonal antibodies against two Golgi proteins. We show that the monoclonal antibodies are highly specific and sufficiently sensitive to detect endogenous proteins in crude extracts by immunoblotting with little background staining. By immunofluorescence the major compartments of the membrane traffic system (including the endoplasmic reticulum, the Golgi, and early and late endosomes) are labeled by at least one antibody. Moreover, the antibodies can be used to label organelles in fly tissues including salivary glands and wing imaginal discs. We anticipate that these antibodies will provide a useful tool kit to facilitate the investigation of how the endomembrane system functions and varies in the diverse tissue types of metazoans. |
format | Online Article Text |
id | pubmed-4958275 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The Company of Biologists Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-49582752016-08-04 An antibody toolkit for the study of membrane traffic in Drosophila melanogaster Riedel, Falko Gillingham, Alison K. Rosa-Ferreira, Cláudia Galindo, Antonio Munro, Sean Biol Open Methods & Techniques The use of Drosophila melanogaster as a model organism has been pivotal to understanding the developmental processes of metazoans. However, the use of flies for studying subcellular organization is hampered by a paucity of reliable reagents to label specific organelles. Here, we describe the generation of mouse monoclonal antibodies against a set of markers of the secretory and endocytic pathways, along with goat polyclonal antibodies against two Golgi proteins. We show that the monoclonal antibodies are highly specific and sufficiently sensitive to detect endogenous proteins in crude extracts by immunoblotting with little background staining. By immunofluorescence the major compartments of the membrane traffic system (including the endoplasmic reticulum, the Golgi, and early and late endosomes) are labeled by at least one antibody. Moreover, the antibodies can be used to label organelles in fly tissues including salivary glands and wing imaginal discs. We anticipate that these antibodies will provide a useful tool kit to facilitate the investigation of how the endomembrane system functions and varies in the diverse tissue types of metazoans. The Company of Biologists Ltd 2016-06-02 /pmc/articles/PMC4958275/ /pubmed/27256406 http://dx.doi.org/10.1242/bio.018937 Text en © 2016. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
spellingShingle | Methods & Techniques Riedel, Falko Gillingham, Alison K. Rosa-Ferreira, Cláudia Galindo, Antonio Munro, Sean An antibody toolkit for the study of membrane traffic in Drosophila melanogaster |
title | An antibody toolkit for the study of membrane traffic in Drosophila melanogaster |
title_full | An antibody toolkit for the study of membrane traffic in Drosophila melanogaster |
title_fullStr | An antibody toolkit for the study of membrane traffic in Drosophila melanogaster |
title_full_unstemmed | An antibody toolkit for the study of membrane traffic in Drosophila melanogaster |
title_short | An antibody toolkit for the study of membrane traffic in Drosophila melanogaster |
title_sort | antibody toolkit for the study of membrane traffic in drosophila melanogaster |
topic | Methods & Techniques |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4958275/ https://www.ncbi.nlm.nih.gov/pubmed/27256406 http://dx.doi.org/10.1242/bio.018937 |
work_keys_str_mv | AT riedelfalko anantibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT gillinghamalisonk anantibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT rosaferreiraclaudia anantibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT galindoantonio anantibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT munrosean anantibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT riedelfalko antibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT gillinghamalisonk antibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT rosaferreiraclaudia antibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT galindoantonio antibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster AT munrosean antibodytoolkitforthestudyofmembranetrafficindrosophilamelanogaster |