Apoptosis assay cell type - RAW 264.7 mouse macrophage

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 2 matching solutions for this experiment

Upstream tips
- Prepare 1x Binding Buffer by diluting 1 ml of the 10x Binding Buffer with 9 ml of deionized water.

- Dissolve the staurosporine in DMSO to a concentration of 100 mg/ml.

- cells were pretreated with BER for 1 h, then cultured with or without fMLP (1µM) stimulation for 12 h
Downstream tips
- Apoptotic neutrophils are here defined as Annexin V-positive but propidium iodide-negative cells.
Protocol tips
Add dyes and incubate for 30 min, at 37°C/5% CO2
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