HK-2 cells (1×107 cells/well) were seeded into 6-well plates, and SchB or cis-DDP was added 24 h later. Following the appropriate incubation period, the cells were stained with Annexin V-fluorescein isothiocyanate and propidium iodide (Beyotime Institute of Biotechnology) for 30 min. A flow cytometer (BD FACSVerse; BD Biosciences, Franklin Lakes, NJ, USA) was used to determine the apoptotic rate using CellQuest Pro 3.3 software (BD Biosciences).
Cellquest pro 3
Manufactured by BD
Sourced in United States
CellQuest Pro 3.3 software is a flow cytometry analysis tool developed by BD. It provides data acquisition, analysis, and reporting capabilities for flow cytometry experiments.
Automatically generated - may contain errors
Lab products found in correlation
Facsverse, by BD (1 mentions)
Propidium iodide, by Beyotime (1 mentions)
Annexin 5 fluorescein isothiocyanate, by Beyotime (1 mentions)
Facscalibur, by BD (1 mentions)
2 7 dichlorofluorescein dcf diacetate, by Thermo Fisher Scientific (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Annexin 5 apc 7 aad apoptosis detection kit, by Keygen Biotech (1 mentions)
4 protocols using cellquest pro 3
1
Apoptosis Assay in HK-2 Cells
2
Intracellular ROS Measurement in BALF Cells
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ROS were measured as previously described (22 (link)). BALF cells were washed with PBS. To measure intracellular ROS, cells were incubated for 10 min at room temperature with PBS containing 3.3 µM 2′,7′-dichlorofluorescein (DCF) diacetate (Molecular Probes, Eugene, OR, USA), to label intracellular ROS. DCF-stained cells (1×104) were subjected to fluorescence-activated cell sorting analysis to measure ROS levels using a FACSCalibur instrument (BD Biosciences, San Jose, CA, USA). The data were analyzed with CellQuest Pro 3.3 software (BD Biosciences).
3
Apoptosis Quantification via Flow Cytometry
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Following 24 h of transfection, the cells were seeded into six-well plates at 1×105 cells/well. The cells from each group were collected following drug treatment and washed twice with PBS. The cells were then re-suspended in 500 µl binding buffer and mixed sequentially with 5 µl Annexin V-allophycocyanin (APC) and 5 µl 7-aminoactinomycin D (7-AAD) using an Annexin V-APC/7-AAD Apoptosis Detection kit (cat no. KGA1025; Nanjing KeyGen Biotech Co., Ltd., Nanjing, China), according to the manufacturer's instructions. The cells were subsequently incubated at room temperature in the dark for 15 min, and apoptotic rates were detected using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, USA). The data were analyzed using CellQuest Pro 3.3 software (BD Biosciences).
4
Annexin V-FITC Apoptosis Assay
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The cells from each group were collected following trypsin free of EDTA treatment gently and were centrifuged at 2000 rpm for 5 min and washed twice with PBS. The cells were re-suspended with the binding buffer and 100 μl (a final concentration of 106 cells/ml) suspended cells were added to negative control tube one, negative control two and positive control tube one, positive control tube two. Then, when all of tubes were mixed to 5 μl Annexin V-FITC staining solution, the cells were subsequently incubated at 2–8 °C in the dark for 5 min and the apoptotic rates were detected using a flow cytometer (BD Biosciences, Franklin Lakes, USA). The data were analyzed using CellQuest Pro 3.3 software (BD Biosciences).
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