The H9c2 cell culture medium was sucked out into a suitable centrifuge tube. Then the adherent H9c2 cells were washed by PBS once and digested by trypsin cell digestion solution, which contained EDTA (Solarbio, T1300-100, Beijing, China). After incubation at room temperature, we added the cell culture medium which was collected before, and transfered it to the centrifugal tube, then centrifuged 1000 g for 5 min (Bioridge, TDZ4-WS, Shanghai, China). The H9c2 cells were centrifuged and resuspended with 195 μl Annexin V-FITC binding solution and 5 μl Annexin V-FITC (Beyotime, C1062, Nantong, China), incubated at 4°C for 15 min. Then, the cells were centrifuged again and resuspended with 190 μl Annexin V-FITC binding solution and 10 μl propidium iodide staining solution (Beyotime, Nantong, China), incubated at 4°C for 5 min. The fluorescence was detected by flow cytometer (BD, Accuri C6, United States).
Propidium iodide staining solution
Manufactured by Beyotime
Sourced in China, United States
Propidium iodide staining solution is a nucleic acid-binding dye used in flow cytometry and fluorescence microscopy applications. It selectively binds to the DNA of cells, allowing for the identification and quantification of dead or dying cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (4 mentions)
Annexin 5 fitc staining solution, by Beyotime (4 mentions)
Annexin 5 fitc, by Beyotime (3 mentions)
Accuri c6, by BD (3 mentions)
Annexin 5 fitc binding solution, by Beyotime (2 mentions)
Flow cytometry, by BD (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Facscan, by BD (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
T1300 100, by Solarbio (1 mentions)
C1062, by Beyotime (1 mentions)
Annexin 5 fifc, by Beyotime (1 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
Version 10, by FlowJo (1 mentions)
Facscanto 2, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Cell cycle and apoptosis detection kit, by Beyotime (1 mentions)
Binding buffer, by Beyotime (1 mentions)
Facs caliber system, by BD (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
32 protocols using propidium iodide staining solution
1
H9c2 Cell Apoptosis Assay
2
Quantifying Apoptosis in Ovarian Cancer Cells
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Epithelial ovarian cancers cells were collected into a centrifuge tube and washed with the precooled phosphate buffer saline (PBS). Then, 70% ethanol that was precooled in an ice bath was added and gently mixed at 4°C for 2 hours or more. Next, precooled PBS was used to wash the cells. Each tube containing cell samples were supplemented with 0.5 mL of propidium iodide staining solution (Beyotime, C1052), the cell precipitation was slowly and fully resuscitated, and the solutions were incubated at 37°C for 30 minutes in the dark. Flow cytometry was used to detect red fluorescence at the excitation wavelength of 488 nm.
3
Cell Cycle Analysis of UV-Irradiated HL-60 Cells
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HL-60 cells were plated in a 24-well plate at a density of 1×106 cells/well and exposed to UV LED irradiation at 0, 8, 15 and 30 J/m2. Following incubation for 2 h, cells were harvested and resuspended in PBS and fixed in 70% ethanol at 4°C overnight. They were then washed twice in cold PBS and incubated with propidium iodide staining solution (Beyotime Institute of Biotechnology, Haimen, China) for 30 min at room temperature. The percentage of cells at various phases of the cell cycle, namely the G0/G1, S and G2/M phases, were determined by flow cytometric analysis of 1×105 cells.
4
Apoptosis Assay by Flow Cytometry
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Suspended and adherent cells were collected and washed with cold PBS. Up to 500 μl of binding buffer was added to resuspend the cells. Annexin V-FIFC (5 μl) and propidium iodide staining solution (5 μl; Beyotime, Jiangsu, China) were added for 20 min. and incubated in the dark. The apoptotic rate was measured by flow cytometry (Beckman, Fullerton, CA, USA).
5
Cell Cycle Analysis of HSPCs
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HSPCs (106) were washed with cold PBS and fixed in cold 4% paraformaldehyde for 1 h at room temperature. Cells were centrifuged at 1000 g for 5 min, washed with PBS, and fixed overnight in 70% ethanol at 4°C. Cells were then centrifuged at 1000g for 5 min, washed with PBS, and incubated in propidium iodide staining solution (Beyotime) at 37°C for 30 min in the dark. Flow cytometry was performed using an excitation wavelength of 488 nm. The cell cycle distribution was analyzed using FACS Express software.
6
Apoptosis Assessment by Flow Cytometry
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The neurons were collected and the cells were rinsed with cold PBS buffer. After that, the cells were followingly resuspended in binding buffer. In each sample, 5 μl of Annexin V‐FITC staining solution and 5 μl of propidium iodide staining solution (Beyotime, Shanghai, China) were loaded, and then incubated at 4°C protected from light and stained for 30 min. The cells were subsequently rinsed three times with the binding buffer to remove excessive dye, and then resuspended in 500 μl of binding buffer. Then, the cells were detected with a BD FACSCanto II flow cytometer (BD Biosciences, San Jose, CA, USA) within 1 h, and the data were followingly analyzed by a FlowJo version 10.2 software (FlowJo, LLC Ltd, Ashland, OR, USA).
7
Cell Cycle Analysis by Flow Cytometry
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Cell cycle analysis was performed using flow cytometry. BC cells were collected and fixed in 70% ethanol. The cells were then incubated with a propidium iodide staining solution (Beyotime, China) for 30 min in the dark. Samples were analyzed using a FACS Canto II flow cytometer (BD Bioscience, USA).
8
Cell Cycle Analysis by Flow Cytometry
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Flow cytometry was used to detect the proportion of cells in each cell cycle phase using the Cell Cycle and Apoptosis Detection kit (Beyotime) according to the manufacturer’s instructions. ICP1 cells were transfected with plasmid DNA (pCMV-HA-
CDKN3, pCMV-HA) or siRNA (si-
CDKN3 or si-NC). Forty-eight hours later, the cells were harvested and fixed in 70% ethanol at –20°C overnight. After removal of the fixation medium by centrifugation at 266
g, 500 μg of propidium iodide staining solution (Beyotime) was added to the pelleted cells. The mixture was incubated at 37°C for 30 min. Red fluorescence was observed with a flow cytometer (Beckman Coulter, Brea, USA) at an excitation wavelength of 488 nm.
CDKN3, pCMV-HA) or siRNA (si-
CDKN3 or si-NC). Forty-eight hours later, the cells were harvested and fixed in 70% ethanol at –20°C overnight. After removal of the fixation medium by centrifugation at 266
g, 500 μg of propidium iodide staining solution (Beyotime) was added to the pelleted cells. The mixture was incubated at 37°C for 30 min. Red fluorescence was observed with a flow cytometer (Beckman Coulter, Brea, USA) at an excitation wavelength of 488 nm.
9
Cell Cycle Analysis by Flow Cytometry
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At 24 or 48 h after seeding, JB6 cell pellets were collected following treatment with 0.25% trypsin. The cells were washed with prechilled PBS, fixed with 70% ethanol and stained with propidium iodide staining solution (Beyotime, China) at 37 °C for 30 min in the dark. Red fluorescence was detected by flow cytometry. FlowJo 7.0 software was used for DNA content and light scattering analysis.
10
Apoptosis Assessment in H1299 and H1650 Cells
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H1299 and H1650 cells were resuspended in binding buffer (Beyotime Institute of Biotechnology) and cell concentration was adjusted to 1×106 cells/ml. In each group, 100 µl cell suspension was mixed with 5 µl annexin V/FITC staining solution and 10 µl propidium iodide staining solution (20 µg/ml; Beyotime Institute of Biotechnology), followed by incubation at room temperature for 15 min in the dark. Next, a flow cytometer (FACScan; BD Biosciences) was used to examine cell apoptosis and the results were analyzed using FlowJo (version 10; FlowJo LLC).
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