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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
The Company of Biologists
2016-07-01
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| Series: | Biology Open |
| Subjects: | |
| Online Access: | http://bio.biologists.org/content/5/7/987 |
| _version_ | 1818621052791554048 |
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| author | Falko Riedel Alison K. Gillingham Cláudia Rosa-Ferreira Antonio Galindo Sean Munro |
| author_facet | Falko Riedel Alison K. Gillingham Cláudia Rosa-Ferreira Antonio Galindo Sean Munro |
| author_sort | Falko Riedel |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-16T18:03:09Z |
| format | Article |
| id | doaj.art-223eec48bea942b0ac6c6ce473cc98d3 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-16T18:03:09Z |
| publishDate | 2016-07-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-223eec48bea942b0ac6c6ce473cc98d32022-12-21T22:22:00ZengThe Company of BiologistsBiology Open2046-63902016-07-015798799210.1242/bio.018937018937An antibody toolkit for the study of membrane traffic in Drosophila melanogasterFalko Riedel0Alison K. Gillingham1Cláudia Rosa-Ferreira2Antonio Galindo3Sean Munro4 MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH, UK MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH, UK MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH, UK MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH, UK MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH, UK 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.http://bio.biologists.org/content/5/7/987Organelle markersDrosophilaGolgiEndosomesEndoplasmic reticulum |
| spellingShingle | Falko Riedel Alison K. Gillingham Cláudia Rosa-Ferreira Antonio Galindo Sean Munro An antibody toolkit for the study of membrane traffic in Drosophila melanogaster Biology Open Organelle markers Drosophila Golgi Endosomes Endoplasmic reticulum |
| 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 | Organelle markers Drosophila Golgi Endosomes Endoplasmic reticulum |
| url | http://bio.biologists.org/content/5/7/987 |
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