BioLegend
Alexa Fluor® 647 Annexin V
640911
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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.
Flow Cytometry
Cultured B cells treated with an agonistic anti-Fas antibody or isotype control for 4 hours at 37 deg C
Annexin V (1:20) in Annexin V Binding Buffer (BioLegend), 15 min at room temp.
N/A
Dilute cells 5-fold with Annexin V Binding Buffer and analyze by flow cytometry
Flow cytometry with a 633 nm laser
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.
Clean discrimination of apoptotic and non-apoptotic cells; does not require cell fixation
None
We'll continue to use this reagent for apoptosis measurements.