An antibody toolkit for the study of membrane traffic in Drosophila melanogaster

被引:59
作者
Riedel, Falko [1 ]
Gillingham, Alison K. [1 ]
Rosa-Ferreira, Claudia [1 ]
Galindo, Antonio [1 ]
Munro, Sean [1 ]
机构
[1] MRC Lab Mol Biol, Cambridge Biomed Campus,Francis Crick Ave, Cambridge CB2 0QH, England
基金
英国惠康基金; 英国医学研究理事会;
关键词
Organelle markers; Drosophila; Golgi; Endosomes; Endoplasmic reticulum; SECRETORY PATHWAY; RAB PROTEINS; TER SITES; BIOGENESIS; NOTCH; ORGANIZATION; ACTIVATION; LYSOSOMES; FUSION; CELLS;
D O I
10.1242/bio.018937
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:987 / 992
页数:6
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