Clear Resolution Of Live Apoptotic And Non-Apoptotic Cells

Immunology
Duke University
Graduate student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Alexa Fluor® 647 Annexin V

Catalog Number:

640911

The aim was to study the role of various factors in B-cell survival in vitro. Annexin V staining was used to detect apoptotic cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Cultured B cells treated with an agonistic anti-Fas antibody or isotype control for 4 hours at 37 deg C

Primary Incubation

Annexin V (1:20) in Annexin V Binding Buffer (BioLegend), 15 min at room temp.

Blocking Agent

N/A

Secondary Incubation

Dilute cells 5-fold with Annexin V Binding Buffer and analyze by flow cytometry

Tertiary Incubation

N/A

Detection

Flow cytometry with a 633 nm laser

Results Summary

Treatment with an isotype control antibody (Hamster IgG, BioXCell) resulted in ~5% apoptotic B cells. Treatment with anti-Fas antibody (clone Jo2, BD Biosciences) resulted in >80% apoptotic B cells.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Clean discrimination of apoptotic and non-apoptotic cells; does not require cell fixation

The Bad

None

The Bottom Line

We'll continue to use this reagent for apoptosis measurements.

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