Annexin 5 biotin apoptosis detection kit

The state of apoptosis was analyzed by Annexin V-biotin apoptosis detection kit (BioVision, Milpitas, CA, USA) following double staining with fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI). Annexin V binds to apoptotic cells with exposed phosphatidylserine, while PI binds to the late apoptotic and necrotic cells with membrane damage. ESCC cells were treated for 1, 6, 12, and 48 h. Staining was performed according to the manufacturer’s instructions.

Immunofluorescence staining of cultured cardiac myocytes and myocardial tissues was performed using Tyramide Signal Amplification (TSA) technology for fluorescence (TSA^TM Biotin System, PerkinElmer). Double-immunostaining for cardiac myosin was performed as described elsewhere9 (link). To stain for AIF, the cells were fixed with 4% paraformaldehyde in PBS for 15 min and then incubated in the rabbit anti-AIF mAb (E20; Epitomics Inc.). Confocal microscopy was performed using an LSM510 laser scanning microscope (Zeiss). To stain for Annexin-V, the cells were incubated in biotinylated Annexin-V in 1X binding buffer (Annexin V-Biotin Apoptosis Detection Kit, BioVision Inc.) for 5 min and then fixed with 2% paraformaldehyde in PBS for 15 min.

Measurement of the percent of apoptotic cells was performed using an Annexin V-Biotin Apoptosis Detection kit (BioVision Inc., Milpitas, CA, USA) per the manufacturer’s instructions. Cells were analyzed using a FACSAria™ Cell Sorter.