Overview
Apoptosis, or programmed cell death, is an important regulator of cell growth and proliferation. Breakdowns in the apoptotic pathway can also lead to cancer and various diseases.Combining the power of Guava flow cytometry systems and the assay development expertise of Chemicon and Upstate, Millipore offers multiple Guava assays for the detection of early-, mid-, and late-stage apoptosis. These kits analyze characteristic physiological changes, including translocation of phosphatidylserine (PS) to the cell surface, cleavage and degradation of specific cellular proteins, compaction and fragmentation of nuclear chromatin, and loss of membrane integrity.
Optimized for use on Guava Systems and requiring only micro-volume cell samples, these assays provide superior discrimination between apoptotic and non-apoptotic cells, as well as detailed profiles of populations in early, mid, and late stages of apoptosis.
Early Apoptosis
Changes in the cellular membrane are among the hallmarks of the early stages of apoptosis. Phosphatidyl serine (PS), normally located inside the cell membrane, moves to the outer surface, and mitochondrial membrane potential drops.Millipore offers two kits that analyze these changes:
- The Guava Nexin kit, which utilizes the binding of annexin V to PS residues for detection
- Guava Mitochondrial Depolarization Assay, which monitors the loss of membrane potential.
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Mid-Stage Apoptosis
Caspases are involved in propagating mid-stage apoptotic signaling cascades, carrying out cellular breakdown, and processing cytokines. Kinetics and dose response curves for specific caspase activation can provide valuable information about their unique properties and mechanisms of action.Millipore’s suite of Guava Caspase Assays employ optimized FLICA (fluorescent labeled inhibitor of caspases) reagents to easily discriminate between the stages of the apoptotic process. Kits are available for caspase 3/7, 8, 9, and multiple caspases.
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Late Apoptosis
One of the hallmarks of late-stage apoptosis the fragmentation of nuclear chromatin, which yields DNA strands with exposed 3'-hydroxyl ends.The Guava TUNEL Assay applies the classic TUNEL (terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling) method to flow cytometry, enabling the detection of apoptosis-induced DNA fragmentation through a quantitative fluorescence assay.
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