Journal
/
Biochemistry
/
Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry
JoVE Journal
Biochemistry
Author Produced
This content is Free Access.
JoVE Journal Biochemistry
Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

DOI:
10.3791/64745-v

•

05:29 min

•

March 24, 2023

•

, ,
1Institute of Experimental Immunology,University of Zürich

Chapters

  • 00:04Introduction
  • 00:45Seeding, Cell Treatment, and Preparation of Cell Lysate
  • 02:10BCA Assay for Protein Quantification
  • 02:49Population-Based Assay for Caspase-3/7 Activity
  • 03:31Single-Cell Assay for Caspase-3/7 Activity
  • 04:07Results: Smac Mimetic-Induced Effect on Caspase-3/7 Activity
  • 04:50Conclusion

Summary

Automatic Translation

Automatic Translation

The present protocol describes two methods to measure caspase activity through a fluorogenic substrate using flow cytometry or a spectrofluorometer.

Tags

Caspase ActivityFluorometric AssayFlow CytometryCell DeathSMAC MimeticBone Marrow-derived Macrophages (BMDMs)DISC Lysis BufferBCA AssayProtein QuantificationFluorometric Instrument

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

A Fluorescence-based Assay of Phospholipid Scramblase Activity

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

A Fluorogenic Peptide Cleavage Assay to Screen for Proteolytic Activity: Applications for coronavirus spike protein activation

A Colorimetric Assay of Citrate Synthase Activity in Drosophila Melanogaster

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Read Article