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Protocol to study internalization and localization dynamics of exogenously added proteins in detergent-permeabilized
Sritama Ray1, Kamalika Mukherjee2, Suvendra N Bhattacharyya3
1CSIR-Indian Institute of Chemical Biology, Kolkata, West Bengal 700032, India.
STAR Protocols
|July 31, 2025
Summary
This study presents a protocol for tracking exogenous proteins within detergent-permeabilized human cells. The method visualizes protein internalization and subcellular localization using immunofluorescence, aiding functional studies.
Area of Science:
- Cell biology
- Biochemistry
- Molecular biology
Background:
- Cell-permeabilization and resealing assays are crucial for investigating exogenous factors in cells.
- Understanding the localization and function of introduced proteins is vital in biological research.
Purpose of the Study:
- To describe a detailed protocol for studying the internalization and subcellular localization of exogenous proteins in detergent-permeabilized human cells.
- To provide a method adaptable for various experimental conditions and research goals.
Main Methods:
- The protocol involves culturing human cells, permeabilization, and preparation of ghost cells.
- Lysate incubation, fixation, and subsequent immunofluorescence staining and imaging are employed.
- The method focuses on visualizing subcellular protein localization.
Main Results:
- The described protocol enables the visualization of exogenous protein internalization and localization.
- Immunofluorescence staining and imaging confirm the subcellular distribution of introduced proteins.
Conclusions:
- The developed assay is an effective tool for studying exogenous protein behavior in permeabilized cells.
- This protocol can be adapted for diverse research applications in cell biology and related fields.
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