Apoptosis Assays
An apoptosis assay detects and quantifies the cellular events associated with programmed cell death, including caspase activation, cell surface exposure of phosphatidylserine (PS) and DNA fragmentation.
Promega apoptosis assays include the luminescent Caspase-Glo® Assays, which detect activity of caspases involved in either the intrinsic or extrinsic apoptosis pathways. The Caspase-Glo® 3/7, 8 and 9 Assays are the most sensitive caspase assays available.
The RealTime-Glo® Annexin V Apoptosis Assay is a luminescent assay that detects phosphatidylserine (PS) exposure and continually monitors apoptosis in real time within a cell culture well.
We also offer TUNEL assays, apoptosis antibodies, inhibitors and other markers for apoptosis.
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Apoptosis Assay Basics
Apoptosis, or programmed cell death, eliminates damaged, infected or redundant cells. Apoptosis is associated with distinct morphological changes, including cell shrinkage, chromatin condensation, loss of nuclear membrane integrity, plasma membrane blebbing, and the formation of apoptotic bodies. Other types of cell death include necrosis, which occurs after cellular injury and is less regulated than apoptosis. Understanding if cell death is a result of apoptosis is important for understanding developmental pathways, the mechanism of action of small molecule and large molecule (biologics) drugs, and related functions of the immune system.
Apoptosis assays typically detect biochemical and cellular events that indicate activation of an apoptotic pathway. The activation of an effector (e.g., caspase-3) leads to common apoptotic events such as exposure of phosphatidylserine on the outer surface of the plasma membrane (typically measured through annexin V binding), cleavage of cellular homeostatic and repair enzymes (e.g., poly [ADP-ribose] polymerase [PARP] measured by antibody techniques) and internucleosomal DNA fragmentation (typically measured through TUNEL assays). Caspase-3 activation is considered full commitment to apoptotic cell death. Exactly which events occur will depend on the apoptotic pathway activated.
Typically, more than one method is necessary to confirm that cell death is occurring via apoptosis. Markers of apoptosis such as caspase activity may be expressed only transiently. Therefore, to determine if apoptosis is the primary mechanism of cell death, understanding the kinetics of the cell death process in your model system is critical. Real-time, live-cell assays monitor apoptotic events over time. Multiple methods may also be used to assess the timing of events within the apoptotic pathway.
