Apoptotic & Necrotic Cell Differentiation

• Convenient simultaneous detection apoptotic and necrotic - as well as healthy cells - in a single assay
• Fast, one-step procedures - assays completed within 15 minutes
• Optimized sensitivity 

Apoptosis and necrosis are the two major processes leading to cell death. Apoptosis is an active, genetically regulated "cell suicide" induced by diverse factors. This disassembly of the cell creates multiple events and is associated e.g. with changes in the phospholipid content of the outer leaflet of the cytoplasmic membrane. Phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition.
Thus, apoptotic cells can be easily and reliably identified with Annexin V, a phospholipid protein with a high affinity for PS, that binds specifically to the PS exposed on the outer membrane leaflet of apoptotic cells. When Annexin V is labeled e.g. with fluorescein (FITC) apoptotic cells are stained brightly green. 
Necrosis normally results from a severe cellular insult. Since both internal organelle and plasma membrane integrity are lost, cytosolic and organellar contents are released into the surrounding environment. Ethidium homodimer III (EtD-III) is a nucleic acid probe, which is impermeant to live or apoptotic cells, but stains necrotic cells intensively with red fluorescence. Due to its significantly higher affinity for DNA and higher fluorescence quantum yield, EthD-III is a superior alternative to propidium iodide (PI) or ethidium homodimer I (EthD-I) commonly used in some other cell staining kits.

The PromoKine Apoptotic/Necrotic Cells Detection Kit provides a convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy.

The PromoKine Apoptotic/Necrotic/Healthy Cells Detection Kit additionally includes a membrane-permeant blue fluorescent DNA stain (Hoechst 33342) for quantifying the healthy cell population. Thus, this kit allows simultaneous quantification of apoptotic (green), necrotic (red) and healthy cells (blue) within the same cell population by flow cytometry or fluorescence microscopy.

Figure:
Fluorescent staining of apoptotic (green), necrotic (red) and viable (blue) cells using PromoKine's Apoptotic/Necrotic/Healthy Cells Detection Kit.