Apoptosis assay cell type - HT-29 human colon adenocarcinoma

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 3 matching solutions for this experiment

Upstream tips
- 5 × 10^4 cells/well were seeded.

- 24h post seeding, cells were treated with MINS for 24 h, 36 h or 48 h.
Protocol tips
- First cells were stained with propidium iodide (50 μg/mL) in the dark for 60 min at 4˚C.

- Next stained with Annexin V-FITC in the dark for 15 min at room temperature.

- Lastly, cells were stained with Hoechst 33342 (10 µg/mL)
in the dark for 15 min at room temperature
Upstream tips
- Cells were pretreated with CM and CF at concentrations required for 50% inhibition of growth
Protocol tips
- Always stain cells with Annexin before fixation (makes cell membrane leaky)
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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