Tunel assay kit cat no g3250

After anaesthesia, a separate group of mouse hearts underwent rapid excision and fixation by 4% paraformaldehyde solution for 12 h at 4°C. Following dehydration by a vacuum tissue processor (Thermo Fischer Scientific), hearts from different groups were embedded by paraffin and cut into 3–4 sections. To detect apoptosis, a commercial TUNEL assay kit (Cat No. G3250) from Promega (Madison, WI, USA) was employed following the instructions of the manufacturer. After the TUNEL and immunofluorescence staining for the myocardium marker cTnT, a LSM810 confocal microscope (Carl Zeiss AG, Oberkochen, Baden voorburg, Germany) was used to capture corresponding pictures. For quantification, myocardium apoptosis was presented as total nuclei (blue) count devided by TUNEL‐positive nuclei (green) count in every sharp field of each section in AAR region.

The assay was conducted with Promega TUNEL assay kit (cat no. G3250). Myocardial tissue was embedded into paraffin. Then, they were deparaffinized and rehydrated with ethanol. The tissue was fixed in 4% paraformaldehyde and treated with the rTDT incubation buffer. Finally, the sample was counterstained with DAPI and photographs were captured with a fluorescence microscope (Nikon).