Isolation and Staining of Mouse Skin Keratinocytes for Cell Cycle Specific Analysis of Cellular Protein Expression by Mass Cytometry

Overview

This protocol describes a method for isolating skin keratinocytes from mouse models, staining them with metal-tagged antibodies, and analyzing the stained cells using mass cytometry. This approach allows for profiling protein expression patterns across different cell cycle phases.

Key Study Components

Area of Science

• Cell biology
• Neuroscience
• Cancer research

Background

• Understanding cell cycle states is crucial for studying skin cells.
• Previous methods lacked precision in determining cell proliferation phases.
• Analyzing protein expression in various cell cycle stages is essential for research.
• This method enhances the analysis of over 40 different markers.

Purpose of Study

• To determine cell cycle states of isolated skin cells.
• To analyze protein expression patterns in discrete phases of the cell cycle.
• To improve the precision of cell cycle phase determination compared to previous methods.

Methods Used

• Isolation of skin keratinocytes from mouse models.
• Staining with metal-tagged antibodies.
• Analysis of stained cells using mass cytometry.
• Profiling protein expression across different cell cycle phases.

Main Results

• The protocol allows for precise determination of cell cycle phases.
• Enables analysis of over 40 different protein markers.
• Enhances application versatility in cancer research.
• Facilitates understanding of disease-related protein expression patterns.

Conclusions

• This method significantly improves the analysis of skin cell cycle states.
• It provides a robust tool for studying protein expression in various diseases.
• The protocol can be applied in diverse research contexts, including cancer.

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