Reliable Apoptosis Detection Assay

Loyola University Chicago
Microbiology and Immunology
Postdoc fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Promega

Product Name:

Caspase-Glo® 3/7 Assay

Catalog Number:

g8090

Aim: the goal of this experiment is to quantify the caspase 3/7 activity of mouse macrophages infected with viruses.Materials: infected or uninfected cells, pan-caspase inhibitor zVAD, and caspase-glo reagents.Method: follow the manufacture's protocol.Results: virus infection induces caspase 3/7 activity, which is inhibited by zVAD treatment.

Experimental Design and Results Summary

Application

Quantification of apoptotic cell death

Starting Material

Cell lysate from mouse macophages

Protocol Overview

Mouse macrophages were seeded in 24-well plate and infected with virus. After 18 hours infection, medium was removed and 120ul caspase-glo reagent/medium mixture was added to lyse the infected cells at RT for 20min. Follow manufacture's protocol for subsequent steps.

Tips

To save reagents, reducing the recommended amount of reagent won't affect the results relatively

Results Summary

In uninfected cells, the caspase 3/7 activity is low. Both virus 1 and virus 2 infection induce caspase 3/7 activity, but virus 2 is more potent. And caspase 3/7 activity is inhibited by zVAD.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy to use; low variation

The Bad

Not found

The Bottom Line

A good way to quantify caspase 3/7 as an indication of apoptosis. Manufacture's protocol can be modified but won't affect the reliability of result.

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