Twenty-four hours before the transfection, the MCF-7 cell line, MCF-7/DOC and MCF-7/EPI were collected during logarithmic growth phase. After trypsinization and centrifugation, cell suspension was obtained, and seeded in a 24-well plate chamber (medium without antibiotic) at a density of 2×105 cells/well. Cells were grown to 50% confluency and miR-415 mimics or inhibitors were transiently transfected using Lipofectamine 2000 (Invitrogen, USA) according to the specifications of the manufacturer. Six hours after the transfection, the medium containing miRNA-lipo2000 mixture was replaced with fresh complete medium, and the plate was placed in an incubator with 37°C/5% CO2 atmosphere. Cells transfected with empty plasmid served as a control group. The cells were collected for miRNA detection at 24 h after transfection and cells were harvested for protein analyses at 36 h after the transfection. The cell apoptosis assay was performed at 72 h after the transfection using flow cytometry. The Annexin V-APC/PI Apoptosis Detection Kit (Invitrogen, USA) was used for the detection of cell apoptosis. For each group, this experiment was repeated 3 times.
Annexin 5 apc pi apoptosis detection kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Annexin V-APC/PI apoptosis detection kit is a laboratory reagent used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine residues exposed on the surface of apoptotic cells, and propidium iodide (PI), a fluorescent dye that intercalates with the DNA of cells with compromised cell membranes. This combination allows for the identification and discrimination of early apoptotic, late apoptotic, and necrotic cells through flow cytometry analysis.
Automatically generated - may contain errors
Lab products found in correlation
Cytexpert software, by Beckman Coulter (4 mentions)
Cytoflex, by Beckman Coulter (3 mentions)
Cytoflex flow cytometer, by Beckman Coulter (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Transwell plate, by Thermo Fisher Scientific (1 mentions)
Facs analyzer, by BD (1 mentions)
Cytexpert 2, by Beckman Coulter (1 mentions)
7 protocols using annexin 5 apc pi apoptosis detection kit
1
miR-415 Modulation of Breast Cancer Cell Lines
2
Glioma Cell Apoptosis via hucMSCs-sTRAIL
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In this study, the effect of hucMSCs infected with LV-scFv-sTRAIL on U87G apoptosis was investigated using co-culture assay. For co-culture, U87G cells (1 × 105/well) were plated in the lower chamber of the transwell plate (Thermo Fisher Scientific) and incubated for 24 h. Then, hucMSCs (1 × 104/well) in each group were grown in the upper chamber. After another 48 h, glioma cell apoptosis was detected using the Annexin V-APC/PI apoptosis detection kit (Invitrogen, Carlsbad, CA, United States). Briefly, the cells were resuspended in 300 µl binding buffer, followed by the addition of Annexin V-APC solution (5 µl). After 25 min of incubation at 4°C, the cells were resuspended for 10 min in a binding buffer with 5 μl of PI. Apoptotic cells were counted using a FACS analyzer (BD Biosciences).
3
Evaluating Apoptosis Induction in Gastric Cancer
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NCI-N87, SNU16, MKN7, MKN28, and AGS cells were seeded into 6-well plates at a density of 1 × 104 cells/mL, then treated with 10 µM of either LOXO-101, entrectinib, dovitinib, dovitinib lactate, dovitinib dilactic acid, regorafenib, cabozantinib, or crizotinib. Cell death was assessed using an annexin V-APC/PI apoptosis detection kit (Thermo Fisher Scientific, Waltham, MA, USA) with a CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA). The percentages of intact and apoptotic cells were calculated using CytExpert software (version 2.0; Beckman Coulter, INC., Brea, CA, USA).
4
Tepotinib-Induced Cell Death in Gastric Cancer
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The MKN45, SNU620, MKN28, KATO III, and AGS cells were seeded into 6-well plates at a density of 5 × 104 cells/mL and were then treated with 10 nM or 10 µM of tepotinib. Cell death was determined using the annexin V-APC/PI apoptosis detection kit (Thermo Fisher Scientific, Waltham, MA, USA) using a CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA). The percentages of intact and apoptotic cells were calculated using CytExpert software (Beckman Coulter).
5
NTRK Inhibitor-Induced Cell Death
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KM12SM, HT29, and HCT116 cells were seeded into 6-well plates at a density of 5 × 104 cells/mL and then treated with 10 nM of each NTRK inhibitor. Cell death was determined using an annexin V-APC/PI apoptosis detection kit (Thermo Fisher Scientific, Waltham, MA, USA) with a CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA). The percentages of intact and apoptotic cells were calculated using CytExpert software (version 2.0; Beckman Coulter).
6
Combination Drug Cytotoxicity Assay
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AGS, Hs746T, MKN45, SNU620, and SNU638 cells were seeded into six-well plates at 5 × 104/mL and treated with various concentrations of ramucirumab (10 nM) paclitaxel (20 nM), or tepotinib (10 nM) alone or in combination. The concentration ranges were selected by referencing the SNU620 IC50 values (~20 nM paclitaxel; ~10 nM tepotinib). The ramucirumab concentration used was that at which SNU620 cell viability decreased when ramucirumab was combined with paclitaxel. Cell death was quantitated using the Annexin V-APC/PI apoptosis detection kit (Thermo Fisher Scientific) and CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA). The proportions of intact and apoptotic cells were calculated using CytExpert 2.0 software (Beckman Coulter).
7
Foretinib Induces Apoptosis in Gastric Cancer Cells
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SNU620, MKN45, MKN28, and AGS cells seeded into 6-well plates at a density of 5 × 104 cells/mL were treated with foretinib IC50 values. Cell death was determined using the Annexin V-APC/PI Apoptosis Detection Kit (Thermo Fisher Scientific) using a CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA). Percentages of intact and apoptotic cells were calculated using the CytExpert software (Beckman Coulter).
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