( ...)Measuring multiple apoptosis,,,parameters with the Agilent 2100,,,bioanalyzer

A simplified and fast protocol for the analysis of DNA fragmentation during apoptosis
This Application Note describes a simplified and fast protocol for apoptosis DNA fragmentation analysis with the Agilent 2100 bioanalyzer using the DNA LabChip kit. The combination of Annexin V, activecaspase- 3 and DNA laddering analysis on the same platform together with the Cell Fluorescence LabChip kit permits quick confirmation and quantitation of apoptosis in cell populations. A fast and simple protocol was applied for the extraction of fragmented, chromosomal DNA. The increased sensitivity and sizing accuracy of the DNA LabChip kit compared to conventional agarose gel electrophoresis also means that a significant lower amount of apoptotic cells is required for the analysis.
The Agilent 2100 bioanalyzer was introduced by Agilent Technologies as the first commerially available lab-on-a-chip analysis system for the life science laboratory using LabChip products, developed in collaboration with Caliper Technologies Corp. Chip-based approaches for a variety of separation- based techniques have been introduced, addressing DNA, RNA and protein separations.1, 2, 3. Recently the Agilent 2100 bioanalyzer has been extended to enable dual color simple flow cytometric assays4. Apoptosis assays have been demonstrated for quantitative measurements of whole cells5. The combination of cytometric and gel-like separations on the same instrument platform makes it an ideal tool for multi-parametric apoptosis quantitation and confirmation.
Apoptosis
Apoptosis, or programmed cell death, is the outcome of a metabolic cascade that results in cells dying in a controlled manner. It is
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