FITC Annexin V Apoptosis Detection Kit I

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1 × 10^6 cells per well were seeded onto the 100 mm culture dishes. - Treated with various concentrations of bryostatin for 12 h. - It was followed by H2O2 (100 µM) for 48h.	- Treated with FITC Annexin V and PI and incubated for 20 min at RT (25°C) in the dark

Upstream tips
- 1 × 10^6 cells per well were seeded onto the 100 mm culture dishes. - Treated with various concentrations of bryostatin for 12 h. - It was followed by H2O2 (100 µM) for 48h.
Protocol tips
- Treated with FITC Annexin V and PI and incubated for 20 min at RT (25°C) in the dark

Publication protocol

The effect of H2O2 and bryostatin I on induction of apoptosis in Saos-2 osteosarcoma cells
was analyzed using fluorescein isothiocyanate
(FITC)-Annexin V Apoptosis Detection kit I (BD
Biosciences, San Jose, CA, USA). Apoptosis induction was analyzed according to the manufacturer’s instructions. The cells were seeded at a density of 1 × 106 cells per well onto the 100 mm culture dishes and treated with various concentrations of bryostatin for 12 h followed by H2O2 (100 µM) for 48. The control cultures were incubated with H2O2 or DMSO alone.
Following incubation, the cells were harvested and centrifuged for 10 min at 500 × g. The cell pellets obtained were put in Annexin V binding buffer containing 140 mM CaCl2, 10 mM HEPES/NaOH and 2.5 mM MgCl2. To each of the well was then added 5 µl each of FITCconjugated Annexin V and propidium iodide (PI) and incubation was continued under dark conditions for 20 min. Fluorescence-Activated Cell Sorting (FACScan) instrument (Becton-Dickinson, Franklin Lakes, CA, USA) was used for the analysis of the apoptotic cells.

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Download the product protocol from BD Biosciences for FITC Annexin V Apoptosis Detection Kit I below.

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