Summary Defects in protein quality control are the underlying cause of age-related diseases. The western blot analysis of detergent-soluble and insoluble protein fractions has proven useful in identifying interventions that regulate proteostasis. Here, we describe the protocol for such analyses in Drosophila tissues, mouse skeletal muscle, human organoids, and HEK293 cells. We describe key adaptations of this protocol and provide key information that will help modify this protocol for future studies in other tissues and disease models. For complete details on the use and execution of this protocol, please refer to Rai et al. (2021) and Hunt el al. (2021).
Graphical abstract
Highlights • Testing detergent-soluble and insoluble protein fractions estimates proteostasis • Loss of proteostasis increases detergent-insoluble poly-ubiquitinated proteins • Protocol for Drosophila, mouse skeletal muscle, human organoids, and cells • Protocol can be adapted to other tissues and disease models
Defects in protein quality control are the underlying cause of age-related disease. The western blot analysis of detergent-soluble and insoluble protein fractions has proven useful in identifying interventions that regulate proteostasis. Here, we describe the protocol for such analyses in Drosophila tissues, mouse skeletal muscle, human organoids, and HEK293 cells. We describe key adaptations of this protocol and provide key information that will help modify this protocol for future studies in other tissues and disease models.