SUB-immunogold-SEM reveals nanoscale distribution of submembranous epitopes.

ABSTRACT: Electron microscopy paired with immunogold labeling is the most precise tool for protein localization. However, these methods are either cumbersome, resulting in small sample numbers and restricted quantification, or limited to identifying protein epitopes external to the membrane. Here, we introduce SUB-immunogold-SEM, a scanning electron microscopy technique that detects intracellular protein epitopes proximal to the membrane. We identify four critical sample preparation factors contributing to the method's sensitivity. We validate its efficacy through precise localization and high-powered quantification of cytoskeletal and transmembrane protein distribution. We evaluate the capabilities of SUB-immunogold-SEM on cells with highly differentiated apical surfaces: (i) auditory hair cells, r

SUBMITTER: Miller KK 

PROVIDER: S-EPMC11387508 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature