*4.9. Qualitative and Quantitative Analysis of DNA Fragmentation*

Hippocampal CA3 tissues were subject to measurement of the apoptotic DNA fragmentation following status epilepticus [5,26,44,85]. After total DNA was extracted from the hippocampal CA3 tissues, nucleosomal DNA ladders were amplified by a PCR kit for DNA ladder assay (Cat.#: APO-DNA1; Maxim Biotech, San Francisco, CA, USA) to heighten the sensitivity according to the manufacturer's protocol [5,26,44,85]. For quantification of apoptotic DNA fragmentation, a cell death detection ELISA kit (Cat.#11774425001; Roche Molecular Biochemicals) was performed to detect the cytosolic level of histone-associated DNA fragments according to the manufacturer's protocol [5,26,44,85]. Proteins from hippocampal tissues served as the source of antigen, jointly with primary anti-histone antibody and secondary anti-DNA antibody coupled with horseradish peroxidase. The cytosolic nucleosomes were quantitatively measured using 2,2 -azino-di-[3-ethylbenzthiazoline] sulfonate as a substrate. Absorbance was determined at 405 nm and referenced at 490 nm using a Multiskan Spectrum reader (Thermo Scientific, Miami, OK, USA).
