Annexin-V-FLUOS Staining Kit

Apoptosis assay cell type - THP-1

Experiment
Apoptosis assay cell type - THP-1
Product
Annexin-V-FLUOS Staining Kit from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
- 1×10^6 cells were seeded and infected
Protocol tips
- Cells were double stained with Annexin-V-FITC and propidium iodide (PI) at different time points after infection

Publication protocol

Approximately 1×106 NMII infected and unpinfected THP-1 cells were double stained with Annexin-V-FITC and propidium iodide (PI) at different time points post infection by using the Annexin-V-FLUOS Staining Kit according to the manufacturer's protocol (Roche). Following staining, the cells were washed twice with FACS buffer (0.5% BSA) and fixed with 1% paraformaldehyde for 15 min. Fluorescence was detected using a fluorescence-activated cell sorter (FACS Calibur, Becton-Dickinson, Palo Alto, CA) to analyze necrotic (PI+), non-apoptotic (negative for both dyes), early apoptotic (Annexin+/PI−) and late apoptotic cells (Annexin+/PI+). Fluorescence parameters were gated using unstained and single-stained uninfected control cells and 10,000 cells were counted for each sample.

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Papers

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Paper title
Induces Apoptosis during Early Stage Infection via a Caspase-Independent Pathway in Human Monocytic THP-1 Cells
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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Annexin-V-FLUOS Staining Kit below.

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