Apoptosis assay cell type - SMMC-7721, HEPG2 human hepatocellular carcinoma

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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Found 5 matching solutions for this experiment

Upstream tips
- Cells were treated with LfcinB-P13 (range, 0–60 µg/ml) for 24 h.
Protocol tips
- Treated with AV, PI and incubate 10-15 minutes at room temperature.
Downstream tips
- Due to the calcium dependence of the Annexin V:PS interaction, it is critical to avoid buffers containing EDTA or other calcium chelators during
Annexin V experiments.

- Annexin V can only be used as a marker of apoptosis in cells where the plasma membrane is intact because destroying the
integrity of the plasma membrane will allow non-specific binding of Annexin V to PS inside the cell.
Protocol tips
The cell apoptosis was measured by flow cytometry
analysis with an Annexin-V/FITC and propidium iodide
(PI) apoptosis detection kit (BioVision, USA), according to the manufacturer’s standards. Briefly, after 48 h
of transfection, the cells were collected and suspended
in annexin-binding buffer and exposed to Annexin-VFITC and PI for 15 min in the dark at room temperature. The apoptotic percentage of cells was quantified by
flow cytometry
Upstream tips
- Dissolve the staurosporine in DMSO to a concentration
of 100 mg/ml..

- 1 × 10^6 cells/well were cultured in 25 cm2 culture flask for 24h
Protocol tips
- 48h post transfection, cells were treated with Annexin-VFITC and PI for 15 min in the dark at room temperature.
Protocol tips
- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark
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