Protocol to study internalization and localization dynamics of exogenously added proteins in detergent-permeabilized human cells

Summary: Cell-permeabilization and resealing assays are essential tools for studying the localization and functional roles of exogenous factors in detergent-permeabilized cells. We describe a protocol for studying the internalization and localization of exogenous proteins in detergent-permeabilized...

Full description

Saved in:
Bibliographic Details
Main Authors: Sritama Ray, Kamalika Mukherjee, Suvendra N. Bhattacharyya
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725003983
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Summary: Cell-permeabilization and resealing assays are essential tools for studying the localization and functional roles of exogenous factors in detergent-permeabilized cells. We describe a protocol for studying the internalization and localization of exogenous proteins in detergent-permeabilized human cells. It includes detailed steps for culturing, permeabilizing, and preparing ghost cells, followed by lysate incubation and fixation. Subsequent immunofluorescence staining and imaging allow visualization of subcellular protein localization. This assay can be adapted to suit a variety of experimental conditions and research objectives.For complete details on the use and execution of this protocol, please refer to Ray et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667