DNA Fragmentation and Apoptosis Assay
Detection of DNA fragmentation for late apoptosis evaluation using the NC-3000™
During apoptosis, nicks and double-strand breaks in the DNA will cause fragmentation of DNA. This late event of apoptosis is detected using DNA content analysis to measure cell having less than one DNA equivalent (so-called sub-G1 cells)
Key benefits:
- Easy measurement of DNA fragmentation at the single cell level
- Acquisition and analysis in one simple step!
- User friendly protocol with predefined settings
- No RNase treatment required
- No calibration required
- PlotManager for superior data presentation
- Automated PDF reports
- Export of data in FCS/ACS formats
Application Note (.PDF):Overview
During apoptosis, calcium- and magnesium-dependent nucleases are activated which degrade DNA. This means that within the DNA there are nicks and double-strand breaks causing fragmentation. This late event of apoptosis is detected using DNA content analysis to measure cell having less than one DNA equivalent (so-called sub-G1 cells).
Principle
Using fluorescence microscopy and image analysis, the NucleoCounter® NC-3000™ system automates detection of cells with fragmented DNA (sub-G1 cells).
After DAPI staining of fixed cells the sample is analyzed using the NucleoCounter® NC-3000™ system and cellular fluorescence is quantified and apoptotic cells with fragmented DNA are seen as a sub-G1 peak in a DNA content histogram displayed on PC screen.
Markers in the histogram can be used to demarcate apoptotic cells.
Results: Presented in PlotManager
Figure: Jurkat cells were grown in the absence (upper row) or in the presence (lower row) of camptothecin and cells were analysed using the DNA Fragmentation Assay and a NucleoCounter® NC-3000™. Scatter plots and histograms were obtained from the NucleoView™ NC-3000™ software. Markers in the displayed histograms were used to demarcate cells with fragmented DNA (Sub-G1 cells). Colored histogram is a merge between untreated (blue line) and camptothecin treated (red line) samples.
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