Detection of Active Caspases During Apoptosis Using Fluorescent Activity-Based Probes

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Enzymatic activity is crucial for cellular processes.
• Activity-based probes (ABPs) are essential tools for studying enzyme function.
• Caspases play key roles in apoptosis, a critical cellular process.

Purpose of the Study:

• To provide detailed methods for using fluorescent ABPs to detect caspase activity.
• To demonstrate the application of fluorescent ABPs for in vitro biochemical profiling.
• To illustrate the use of fluorescent ABPs for in vivo imaging of protease activity.

Main Methods:

• Utilizing fluorescently labeled activity-based probes (ABPs).
• Biochemical profiling of caspase activity using fluorescent SDS-PAGE.
• In vitro and in vivo imaging of protease activity in live cells, tissues, and animals.

Main Results:

• Established protocols for detecting caspase activity during apoptosis onset.
• Demonstrated successful biochemical profiling of caspase activity.
• Showcased the utility of fluorescent ABPs for live imaging applications.

Conclusions:

• Fluorescent ABPs are versatile tools for studying caspase activity.
• These methods facilitate detailed investigation of apoptosis.
• The described techniques are applicable to both in vitro and in vivo studies.