Necrosis HUVEC

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Found 3 matching solutions for this experiment

Upstream tips
• A live-cell real-time assay that measures the exposure of phosphatidylserine (PS) on the outer leaflet of the cell membrane during the apoptotic process
• Detected by annexin V binding with a simple luminescence signal.
• Assay also includes a cell-impermeant, profluorescent DNA dye, which detects necrosis
Protocol tips
• 25 × 10^3 cells/well on 96 well plates
Downstream tips
• Assay, time-dependent increases in luminescence that occur before increases in fluorescence reflect the apoptotic process.
• A significant time delay between the emergences of PS, indicated by Annexin V binding, leading to a luminescence signal and the loss of membrane integrity visualized by fluorescence signal, indicate an apoptotic phenotype leading to secondary necrosis.
• Increases in fluorescence or increase in both luminescence and fluorescence concurrently consist of necrosis or other non-apoptotic mechanisms.

Acridine Orange - CAS 65-61-2 - Calbiochem + Propidium iodide solution

Upstream tips
For viability determination at each time point, AO/PI double staining was performed. AO is a stain that is permeable to viable cells and can stain the cell’s DNA directly. This dye emits a green fluorescence once it is excited. PI, on the other hand, is a dye that is impermeable to viable cells. It can bind to DNA only when the cells are dead; it emits a red-orange fluorescence instead
Protocol tips
•This kit allows for distinguishing between viable cells, apoptotic cells, and necrotic cells via fluorescence microscopy
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