Annexin 5 in situ cell apoptosis detection kit

As reported [28 (link)], the glioma cell apoptosis and necrosis were determined by the Annexin V In Situ Cell Apoptosis Detection Kit (Roche Molecular Biochemicals, Indianapolis, IN, USA) according to the manufacturer’s protocols. Briefly, after indicated treatment/s, glioma cells were stained with Annexin V and propidium iodide (PI) (Molecular Probes). The cell apoptosis percentage was reflected by Annexin V^+/+/PI^−/− plus Annexin V^+/+/PI^+/+ percentage detected by fluorescence-activated cell sorting (FACS) (BD, Shanghai, China). Annexin V^−/−/PI^+/+ percentage was utilized as an indicator of cell necrosis. The time point for FACS assay was selected based on pre-experiment results.

The Annexin V In Situ Cell Apoptosis Detection Kit (Roche, Indianapolis, IN) [61 , 63 (link)] was employed to test cell apoptosis. In brief, following the applied LB-100 treatment, cells were washed, fixed and stained with Annexin V (2.5 μg/mL) and propidium iodide (PI) (2.5 μg/mL). The cell apoptosis ratio was reflected by Annexin V^+/+/PI^-/- plus Annexin V^+/+/PI^+/+ percentage detected by fluorescence-activated cell sorting (FACS) counter (Beckman Coulter, Shanghai, China).

Following the designated PKC412 treatment, cell apoptosis was tested by the Annexin V In Situ Cell Apoptosis Detection Kit (Roche, Indianapolis, IN) [6 , 10 (link)]. Briefly, cells were washed and stained with Annexin V (Roche) and propidium iodide (PI) (Roche). The cell apoptosis ratio was reflected by Annexin V^+/+/PI^−/− plus Annexin V^+/+/PI^+/+ percentage detected by fluorescence-activated cell sorting (FACS) (Beckman Coulter, Shanghai, China).