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Measuring multiple apoptosis,,,parameters with the Agilent 2100,,,bioanalyzer

was used in conjunction with a 0.2-cm light path ultra-micro cell (10 l volume). The whole spectrum (1901100 nm) was acquired and evaluation was automatically done by the Warburg-Christian method12 with internal wavelength correction at 320/10 nm.
Annexin V apoptosis measurements
A fast annexin protocol as described in reference 13 was used.
Results and Discussion
Besides DNA laddering, cytometric assays like annexin V or caspase analysis are commonly used for apoptosis detection. Figure 1A shows the analysis of annexin V in Jurkat cells on the Agilent 2100 bioanalyzer after 6 hours treatment with 5 M campthotecin. After switching the instrument to electrophoresis mode, extracted DNA from treated Jurkat cells was analyzed using the DNA 12000 LabChip kit. The characteristic DNA laddering further confirmed apoptosis in the sample (figure 1B).


Sizing information, provided by the 2100 bioanalyzer software, shows nucleosomal fragmentation of DNA by evenly distributed peaks in the electropherogram (figure 2). Apoptosis is confirmed in samples by fragmentation of the chromosomal DNA in 180200bp intervals.14

Typical DNA extraction with the described fast protocol leads to 10 L of 2-30ng DNA/L (figure 3). The DNA 12000 concentration range specification is 0.550 ng/L, so no further treatment is required. Band concentration was calculated to be approximately 1 ng/L (1 L loaded on the chip).

Even at low concentrations, apoptotic samples are clearly distinguishable from the negative control and necrotic samples (figure 4, samples 1 to 7). The positive
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