Single-cell western blotting of cytoplasmic cytokeratin 8 proteoforms.

ABSTRACT: Differential detergent fractionation (DDF) enables compartment-specific lysis, offering a strategy to analyze cytoplasmic proteins while preserving the nucleus for downstream assays. However, while this method facilitates sub-cellular resolution, current single-cell approaches using DDF remain limited in their ability to identify proteoforms without compromising nuclear integrity. This limitation is especially pronounced in proteins where their proteoforms present diverse biological functions such as cytokeratin 8 (CK8), a structural protein implicated in several disease states. Here, we present a single-cell western blot (scWB) integrated with DDF to selectively solubilize and separate CK8 proteoforms while preserving nuclear integrity. To evaluate assay applicability and nuclear stabilit

SUBMITTER: Fomitcheva Khartchenko A 

PROVIDER: S-EPMC12466321 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature