Apoptosis Detection with Annexin V/Propidium Iodide

May 16, 2016

Northwestern
Neurological Surgery
Research Fellow

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Company:

eBioScience

Product Name:

Annexin V Apoptosis Detection Kit FITC

Catalog Number:

88-8005-74

While there exists a plethora of methods to detect cellular apoptosis, the Annexin V/Propidium Iodide is a method that has been highly trusted within the literature. Results obtained from this assay indicate number of live cells, cells undergoing early death, and lastly cells that have undergone "late" apoptosis.

Experimental Design and Results Summary

Application

Apoptosis detection

Starting Material

Cultured cells

Protocol Overview

1) Incubate extracellular antibody, if desired2) Multiple washes with wash buffer, then with Annexin buffer3) Incubate cells with Annexin and Propidium Iodide for 10-15 minutes4) Read on a flow cytometer

Tips

Read a sample from each condition sequentially; despite optimal washes, there may be unspecific uptake of Annexin and Propidium Iodide which may skew results.

Results Summary

Double-negative population represents live cells. Annexin (FITC) represents cells that started undergoing apoptosis. Double-positive population (Annexin+Propidium Iodide) represent cells that have already undergone apoptosis (late cell death)

Additional Notes

None

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Summary

The Good

If done correctly, optimal results indicative of apoptosis is obtained. Not too expensive.

The Bad

Difficult to optimize. Despite following protocol, some experiments may be skewed due to unspecific uptake of dyes.

The Bottom Line

Great product for apoptosis detected in cultured cells. It may be difficult to optimize and unspecific uptake of dyes may skew results. Should always run a positive control to verify proper technique.

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