RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay

Necrosis HCT 116

Experiment

Necrosis HCT 116

Product

RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay from Promega

Manufacturer

Promega

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream tips
• Assay measures the real-time exposure of phosphatidylserine (PS) on the outer leaflet of cell membranes during the apoptotic process. • After cells were attached, 2× Sur-X of finally detected concentrations and 2× detection reagents in RealTime-Glo™Annexin V Apoptosis and Necrosis Assay (Promega,JA1011) were mixed in 100μl pre-warming culture medium and the mixture was added to each well.	• The RealTime-Glo™ Annexin V Apoptosis and Necrosis assay is non-lytic and the simple “add-and-read” method allows multiple readings from a single assay well. • The combination and timing of luminescent (annexin V binding) and fluorescent (DNA release) signals is used to differentiate secondary necrosis occurring during late apoptosis from necrosis caused by other cytotoxic events.	• Relative luminescence units (RLU) and relative fluorescence units (RFU) were recorded over time up to 6 h by the spark multimode microplate reader (Tecan Trading AG, Switzerland).

Upstream tips
• Assay measures the real-time exposure of phosphatidylserine (PS) on the outer leaflet of cell membranes during the apoptotic process. • After cells were attached, 2× Sur-X of finally detected concentrations and 2× detection reagents in RealTime-Glo™Annexin V Apoptosis and Necrosis Assay (Promega,JA1011) were mixed in 100μl pre-warming culture medium and the mixture was added to each well.
Protocol tips
• The RealTime-Glo™ Annexin V Apoptosis and Necrosis assay is non-lytic and the simple “add-and-read” method allows multiple readings from a single assay well. • The combination and timing of luminescent (annexin V binding) and fluorescent (DNA release) signals is used to differentiate secondary necrosis occurring during late apoptosis from necrosis caused by other cytotoxic events.
Downstream tips
• Relative luminescence units (RLU) and relative fluorescence units (RFU) were recorded over time up to 6 h by the spark multimode microplate reader (Tecan Trading AG, Switzerland).

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Papers

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Manufacturer protocol

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