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Caspases

Caspases

Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside cells.

The Intrinsic Apoptotic Pathway

The Intrinsic Apoptotic Pathway

Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...

The Extrinsic Apoptotic Pathway

The Extrinsic Apoptotic Pathway

The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...

Apoptosis

Apoptosis

Apoptosis is a combination of two Greek words, 'apo' and 'ptosis,' meaning separation and falling off, respectively. Hippocrates used this word to describe gangrene, which was caused due to bandaging of fractured bones. Apoptosis was distinguished from necrosis in 1970 when John Kerr reported observations of morphological changes occurring during apoptosis. During one experiment, he observed that the disruption of blood supply to the liver tissue resulted in a size reduction of the tissue.

Cellular Injury V: Apoptosis and Autophagy

Cellular Injury V: Apoptosis and Autophagy

Cells respond to damage and stress through highly coordinated processes that decide whether they survive or undergo controlled self-destruction. Two major pathways involved in this regulation are apoptosis, a type of programmed cell death, and autophagy, a survival mechanism that helps cells adapt to adverse conditions.ApoptosisApoptosis removes aged or injured cells to maintain tissue balance. During this process, the cell shrinks, chromatin condenses and fragments, and membrane-bound...

Autophagic Cell Death

Autophagic Cell Death

Christian de Duve discovered “autophagy,” a process in which cellular components are engulfed by membrane-bound organelles called autophagosomes. The autophagosomes then fuse with lysosomes to digest the enclosed contents. Autophagy is generally activated in cells to prevent cell death. However, cell death is triggered when the damage is beyond repair.
Autophagy and Apoptosis
Autophagy can activate apoptosis. In normal conditions, the autophagy activating protein Beclin-1 and pro-apoptotic...

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Related Experiment Video

Updated: Jul 7, 2026

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Published on: March 24, 2023

Apoptosis-associated caspase activation assays.

Scott H Kaufmann¹, Sun-Hee Lee, X Wei Meng

¹Division of Oncology Research, Mayo Clinic College of Medicine, 200 First Street, S.W., Rochester, MN 55905, USA. Kaufmann.Scott@Mayo.edu

Methods (San Diego, Calif.)

|March 4, 2008

Summary

Caspase activation, crucial for apoptosis, can be detected using various methods. This study details assays for measuring caspase activity, substrate cleavage, and conformation changes, highlighting their strengths and limitations.

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Last Updated: Jul 7, 2026

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

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Published on: March 24, 2023

Evaluation of Caspase Activation to Assess Innate Immune Cell Death

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Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Published on: April 16, 2014

Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Caspases are cysteine proteases essential for apoptosis.
They are synthesized as inactive precursors and activated by cellular signals.
Caspase activity is implicated in diverse physiological and pathological processes.

Purpose of the Study:

To describe methods for detecting caspase activation.
To review recent assay improvements.
To outline the strengths and limitations of each detection approach.

Main Methods:

Measurement of caspase catalytic activity.
Immunoblotting to detect substrate cleavage.
Immunolabeling with conformation-sensitive antibodies.
Affinity labeling coupled with flow cytometry or ligand blotting.

Main Results:

Detailed protocols for four distinct caspase activation assays are provided.
Recent advancements enhancing assay sensitivity and specificity are identified.
Comparative analysis of assay methodologies, including their advantages and disadvantages, is presented.

Conclusions:

Multiple validated methods exist for detecting caspase activation.
Assay selection depends on specific research questions and experimental contexts.
Understanding assay limitations is critical for accurate apoptosis research.