For the analysis of the cell cycle, Panc-1 and SW1990 cells were harvested, washed with PBS, stained with binding buffer containing 1 ml DNA staining solution and 10 µl permeabilization solution, and incubated at room temperature for 30 min in the dark according to the manufacturer's instructions (MultiSciences Biotech, Co., Ltd.). For the analysis of cell apoptosis, cells were collected, washed in PBS, and resuspended in a 500 µl mixture of 5 µl Annexin V-FITC (Annexin V-PE) and 10 µl propidium iodide (7-AAD; MultiSciences Biotech, Co., Ltd.). Modfit (Verity Software House, Inc.) and FlowJo V10 (TreeStar, Inc.) software were employed to analyze the cell cycle and apoptosis data, respectively.
7 aad
Manufactured by MultiSciences Biotech
Sourced in China
7-AAD is a fluorescent dye that can be used to stain DNA. It is commonly used in flow cytometry applications to identify and analyze cells.
Automatically generated - may contain errors
Lab products found in correlation
Annexin 5 apc, by MultiSciences Biotech (4 mentions)
Annexin 5 pe, by MultiSciences Biotech (2 mentions)
Tunicamycin, by Solarbio (1 mentions)
Flow cytometry, by BD (1 mentions)
Stf 083010, by MedChemExpress (1 mentions)
Gsk2606414, by Selleck Chemicals (1 mentions)
Z vad fmk, by MedChemExpress (1 mentions)
Tunicamycin, by MedChemExpress (1 mentions)
Edta free trypsin, by Thermo Fisher Scientific (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Binding buffer, by MultiSciences Biotech (1 mentions)
Pi rnase dye, by BD (1 mentions)
Accuri c6 plus, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
12 protocols using 7 aad
1
Cell Cycle and Apoptosis Analysis
2
Apoptosis Assay of Lapatinib-Treated MCF-7 Cells
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The different groups of MCF-7 cells were cultured (1×105 cells/well) and treated with lapatinib (at the IC50 concentration) for 48 h. Cells were then collected, washed with ice-cold PBS, and the cell concentration was adjusted to 5×105 cells/ml using staining buffer. Cell apoptosis was assayed by staining with 5 μl Annexin V-PE and 7-AAD (MultiSciences, China) for 30 min at room temperature in the dark, and the proportion of apoptotic cells was determined with a flow cytometer (Beckman Coulter). Each experiment was performed in triplicate.
3
Apoptosis Analysis of Transfected HLECs
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After lentivirus transfection, HLECs (SRA 01/04) were digested into cell suspensions and adjusted to 1 × 106~3 × 106 cells/mL. The cells were washed with PBS, and 5 μL Annexin V-APC or 10 μL 7-AAD was added following the manufacturer’s protocol (Multisciences Biotech Co., Ltd., Taiwan, China). A flow cytometer (Becton, Dickinson and Company, Lake Franklin, NJ, USA) was used to test immediately after the reaction was completed.
4
Apoptosis Induction in LUAD Cells
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Cells were treated with 10μg/mL of tunicamycin (#T8480, Solarbio, China) and then treated with 1.0 μM GSK2606414 (the PERK inhibitor, Selleck Chemicals, USA), 50 μM STF083010 (the IRE1 inhibitor, MedChemExpress, USA), 20 μM Z-VAD-FMK (apoptotic inhibitor, MedChemExpress, USA), or DMSO for 48 h. tunicamycin, a natural antibiotic, triggers cancer cell apoptosis 14 (link) and has been diffusely applied to the study of tumor cell apoptosis and programmed cell death. All cells were treated with Annexin V-APC and 7-AAD (MultiSciences, China) at room temperature for 30 mins. The percentage of apoptotic LUAD cells was detected by flow cytometry (Becton Dickinson, USA).
5
Cell Viability Analysis by Flow Cytometry
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LoVo-OxR (AC092894.1), HCT116-OxR (AC092894.1), and their control vector cells were seeded into 6-well plates; after 48 h, cells were treated with EDTA-free trypsin (Gibco BRL, Grand Island, NY, USA) for digestion, then washed once with PBS, and incubated with APC and 7-AAD (MultiSciences, Hangzhou, China) for 10 min and analysis was performed by flow cytometry (Beckman Coulter, Fullerton, CA, USA).
6
Apoptosis Detection by Flow Cytometry
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Cells were collected 48 h after transfection and washed once with phosphate buffer saline (PBS). Cell suspension was mixed with 5 μL Annexin V-APC and 10 μL 7-AAD (Multi sciences, China, AP105) for 15 min in a dark room (21 (link)). Then apoptotic cells were detected by flow cytometry (Cytoflex, Beckman Coulter, USA).
7
Apoptosis Assay for Osteosarcoma Cells
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The osteosarcoma cells were inoculated in 6-well plates. After being rinsed with PBS, cells were suspended in 1 × Binding buffer (MultiSciences, China). After Annexin V-PE and 7-AAD were stained (MultiSciences, China), the cell suspension was incubated in the dark circumstance, and then the apoptosis was identified.
8
Cell Cycle and Apoptosis Assessment
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To assess the effect of gene expression on the cell cycle, cells were centrifuged, fixed with ethanol, and washed with PBS; 500 µL of PI/RNase dye (BD Biosciences, USA) was then added. The cells were incubated in darkness for 15 min, and then the cells were analyzed by flow cytometry (BD Accuri C6 Plus; BD Biosciences, USA).
To assess the effect of gene expression on cell apoptosis, cells were centrifuged and resuspended in 50 µL 1× binding buffer. Then, 5 µL of Annexin V-APC and 10 µL of 7-AAD (MultiSciences Biotech, China) were added to the cells, which were then incubated in the dark at 25 °C for 25 min. The cells were immediately analyzed by flow cytometry.
To assess the effect of gene expression on cell apoptosis, cells were centrifuged and resuspended in 50 µL 1× binding buffer. Then, 5 µL of Annexin V-APC and 10 µL of 7-AAD (MultiSciences Biotech, China) were added to the cells, which were then incubated in the dark at 25 °C for 25 min. The cells were immediately analyzed by flow cytometry.
9
Apoptosis Assay with Annexin V
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Cells transfected with lentivirus or exposure to different concentration of cisplatin were harvested with tyrisin without EDTA, washed with PBS for 3 times, resuspended with Annexin binding buffer reaching a concentration of 1 10 6 per 100μl. Then cells were dyed with 5μl Annexin V-APC and 10μl 7-AAD (MultiSciences, Hangzhou, China), incubated with room temperature for 5 minutes. Finally, cells were collected with flow cytometer (FACSCalibur, BD, USA), and analyzed with Flowjo V10 software.
10
Apoptosis Analysis of CFPAC and PANC1 Cells
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The apoptosis of CFPAC and PANC1 cells after indicated treatments was assessed by flow cytometry analysis. CFPAC and PANC1 cells were harvested, washed with PBS, and resuspended in a 500 μl mixture with 5 μl Annexin and 10 μl propidium iodide (7-AAD; MultiSciences Biotech, Co., Ltd., Hangzhou, China). Finally, cell apoptosis rate was detected using a flow cytometer and analyzed with the FlowJo software. The apoptosis rate in each group was calculated by Q2 (late apoptosis) + Q3 (early apoptosis).
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