In this study, we determined the lipid and protein composition of lipid droplets isolated from wild type and the high fat daf-2 mutant C. elegans. We performed RNAi knockdown of the genes encoding the most abundant lipid droplet proteins and found an enrichment of genes required for development, reproduction, and the regulation of fat homeostasis. Furthermore, we confirmed the localization of the acyl-CoA synthetase ACS-4 to the surface of lipid droplets.