Annexin V Apoptosis Assay
Detection of Apoptosis using Annexin V and NC-3000™
Key benefits:
- Information about both early apoptotic and late apoptotic/necrotic cells provided
- Fast automated single cell analysis
- Standardized results – even with different users
- User friendly protocol with predefined settings
- No calibration required
- PlotManager for superior data presentation
- Automated PDF reports
- Export of data in FCS/ACS formats
Application Note (.PDF):Overview
Translocalisation of phosphatidylserine (PS) from the inner to the outer membrane layer happens early in apoptosis and precedes other apoptotic events such as loss of membrane integrity and DNA fragmentation. Annexins are group of cellular proteins that bind to phospholipids in a calcium-dependent manner. A member of this group, Annexin V, has proven to be a useful tool in detecting apoptotic cells since it has a high affinity for PS.
Using fluorescently labeled Annexin V the externalization of PS can easily be detected. Annexin V may also bind to PS on late apoptotic and necrotic cells but as the membrane integrity on these cells has been lost, these cells can be distinguished from early apoptotic cells by the use of the impermeant dye propidium iodide.
Principle
Using fluorescence microscopy and image analysis the NucleoCounter® NC-3000™ system automates detection of apoptotic cells based on phosphatidylserine externalization. Cells are stained with Hoechst 33342, PI and FITC labeled Annexin V.
Hoechst 33342 stains the total cell population, while Annexin V stains apoptotic and necrotic cells. Early apoptotic cells exclude PI, while late stage apoptotic and necrotic cells stain positively for both Annexin V and PI.
Results: Presented in PlotManager
Follow us