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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.

Western Blotting

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

Enzyme-Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.

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Related Experiment Video

Updated: May 9, 2026

Evaluation of Caspase Activation to Assess Innate Immune Cell Death

Evaluation of Caspase Activation to Assess Innate Immune Cell Death

Published on: January 20, 2023

Immunoblotting for active caspase-1.

Christopher Jakobs¹, Eva Bartok, Andrej Kubarenko

¹Institute for Clinical Chemistry and Pharmacology, Unit for Clinical Biochemistry, University Hospital, University of Bonn, Bonn, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|July 16, 2013

Summary

This summary is machine-generated.

This study details robust protocols for detecting inflammasome activation via caspase-1 immunoblotting. These methods enable accurate caspase-1 detection independent of new gene expression, crucial for inflammasome research.

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Last Updated: May 9, 2026

Evaluation of Caspase Activation to Assess Innate Immune Cell Death

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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

Area of Science:

Biochemistry
Molecular Biology
Immunology

Background:

Inflammasome activation is critical in innate immunity and inflammatory diseases.
Caspase-1 detection is the gold standard for assessing inflammasome activity.
Existing methods may be limited by requirements for de novo gene expression.

Purpose of the Study:

To provide standardized protocols for caspase-1 immunoblotting.
To enable inflammasome activation detection independent of gene expression.
To facilitate reproducible research in inflammasome biology.

Main Methods:

Detailed protocols for supernatant sample preparation and precipitation.
Established procedures for polyacrylamide gel electrophoresis (PAGE).
Optimized protein immunoblotting techniques for caspase-1 detection.

Main Results:

Successful implementation of protocols for caspase-1 detection in supernatant samples.
Demonstration of method's independence from de novo gene expression.
Validation of PAGE and immunoblotting for sensitive caspase-1 analysis.

Conclusions:

The presented protocols offer a reliable method for detecting inflammasome activation via caspase-1.
These standardized techniques are valuable for researchers studying inflammatory processes.
The independence from gene expression broadens the applicability of inflammasome research.