Pod apoptosis detection kit

The myocardial tissue (500 mg) was excised from the experimental rats. TUNEL assay was used to detect apoptotic cells under a bright-field microscope by the same expert. Cell number was counted from five different fields from different samples. The detection kit, POD apoptosis detection kit (Roche, USA), was used in accordance with the instructions. The paraffin sections of gastric mucosa were TUNEL staining and maximized 400 times under a microscope.

2-3 months old Wistar male white mice weighing 150–200 g were used after one week for proper acclimatization to the animal house conditions (12 h lighting cycle and 29–31°C temperature) with free access to water and standard rodent chow. All experimental procedures were conducted according to the ethical standards approved by the Institutional Animal Ethics Committee guidelines for animal care and use, Airlangga University, Indonesia. Animals were randomly divided into three groups with 10 animals in each group. The first group was treated as the control group (P0). The second group was treated with cisplatin (20 mg/kg I.p) as a positive control (P1). The third group was injected with glutamine (100 mg/kg, i.v.), a gram glutamine suspended in 10 ml solution of ml 0.9% daily for seven consecutive days, and injected with cisplatin (20 mg/kg I.p) on the seventh day to induce nephrotoxicity (P2). All groups received equivalent volumes of the used vehicles. Mice were sacrificed on the tenth day. The longitudinal section of the left kidney was excised from each animal for immunohistochemical examination.
Glutamine product was purchased from Serva, Germany, cisplatin product from Kalbe Farma, POD apoptosis detection kit (11684817910) from ROCHE, and anti-AIF antibody (AIF monoclonal antibody) from ThermoFisher Scientific (Cat. # MA5-15880).