Cell cycle and cell apoptosis assays were assays by using flow cytometry assays (BD, Franklin Lakes, NJ). The cells were harvested and then washed twice with PBS and resuspended in 100 μL of binding buffer. The cells were fixed in 70% ice‐cold ethanol and after holding overnight at 4 °C, the cells were supplemented with RNaseA (Keygen Biotech) and propidium iodide for 37 °C for 30 minutes. The DNA content of labelled cells was detected using FACS cytometry (BD Biosciences Inc., Franklin Lakes, NJ, USA). Cell apoptosis was assessed by using a flow cytometry assay (BD, Franklin Lakes, NJ). Each experiment was performed in triplicate.
Flow cytometry assay
Manufactured by BD
Sourced in United States
Flow cytometry is an analytical technique that measures and analyzes the physical and chemical characteristics of particles, such as cells, within a fluid as they pass through a laser beam. The core function of a flow cytometry assay is to rapidly analyze and characterize multiple parameters of individual cells or particles in a heterogeneous mixture.
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Lab products found in correlation
Facs cytometry, by BD (1 mentions)
Rnase a, by Keygen Biotech (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Keygen Biotech (1 mentions)
Cellquest 3, by BD (1 mentions)
Annexin 5 fitc pi kit, by BD (1 mentions)
Annexin 5 fitc pi cell apoptosis detection kit, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Fluorescent microscopy, by Zeiss (1 mentions)
Annexin 5 fitc, by Santa Cruz Biotechnology (1 mentions)
Facscan, by Beckman Coulter (1 mentions)
Annexin 5 fitc propidium iodide apoptosis kit, by Beyotime (1 mentions)
Cd29 monoclonal antibody, by Thermo Fisher Scientific (1 mentions)
14 protocols using flow cytometry assay
1
Cell Cycle and Apoptosis Analysis
2
Quantifying Apoptosis by Flow Cytometry
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Cell apoptosis was assessed by using the flow cytometry assay (BD, USA). After treatment, cells were collected by centrifugation, followed by washed twice with cold PBS and suspended in 200 μL binding buffer using the Annexin V-FITC/PI apoptosis detection kit (KeyGEN, China). Afterward, cells were stained with 2 μL Annexin V-FITC and 2 μL PI for 15 min in dark at room temperature. Finally, these cells were analyzed using a flow cytometer (BD, USA) and data were analyzed using the FlowJo software (Treestar, USA).
3
Annexin V-FITC/PI Apoptosis Assay
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After incubation under different conditions for 48 h, PC12 cells were resuspended and stained using a Annexin V-FITC/PI kit (Becton, Dickinson and Company, Franklin Lakes, NJ, U.S.A.) for 20 min in the dark. Apoptotic cells (FITC-positive and PI negative) were distinguished via the flow cytometry assay (BD Biosciences, San Jose, CA, U.S.A.), and the percentage of the apoptotic cells was analyzed using CELL Quest 3.0 software (BD Biosciences).
4
Evaluating Cell Apoptosis via Flow Cytometry
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Cell apoptosis was assessed by using the flow cytometry assay (BD, Franklin Lakes, NJ, USA). HTR-8/SVneo cells were stimulated by AEDPPE or TNF-α for 24 h and re-suspended in 100 μl binding buffer. Cells were then stained with Annexin V-FITC and propidium iodide (PI) for 15 min in the dark. Apoptosis was then measured using flow cytometry.
5
Quantifying Cell Apoptosis by Flow Cytometry
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Cell apoptosis was assessed by flow cytometry assay (BD, FranklinLakes, NJ) [23 ]. In brief, the cells were seeded in 6-well plates (1 × 106 cells/well), followed by 24 h incubation at 37 °C. The cells were then treated with different concentrations of DHM and NDP for 24 h. The assay was performed using the Annexin V-FITC/PI cell apoptosis detection kit (BD Pharmingen, USA) according to the manufacturer’s protocol. Subsequently, the cells were monitored by flow cytometry (FACSCalibur, Becton Dickinson, USA), and the data were analyzed using FlowJo™ software (version 10, FlowJo LLC).
6
EGFP Expression Analysis by Microscopy
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The expression of EGFP
protein was observed by fluorescent microscopy (Zeiss, Germany) with
a 10× objective. The percentage of EGFP-positive cells and mean
fluorescence intensity was quantitatively measured by flow cytometry
assay (BD, USA).
protein was observed by fluorescent microscopy (Zeiss, Germany) with
a 10× objective. The percentage of EGFP-positive cells and mean
fluorescence intensity was quantitatively measured by flow cytometry
assay (BD, USA).
7
Cell Cycle Analysis by Flow Cytometry
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For cell cycle analysis, the cells were harvested and fixed in 70% ice-cold ethanol overnight at 4 °C. The fixed cells were washed twice with phosphate buffered saline (PBS) and resuspended in 0.5 mL PBS containing 0.5 mg/mL RNase and 10 mg/mL PI at 37 °C in the dark for 30 min. The DNA content of the labeled cells was assessed with a flow cytometry assay (BD Biosciences, Franklin Lakes, NJ, USA). Each experiment was performed in triplicate.
8
Apoptosis Analysis by Flow Cytometry
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After transfection, early and late cell apoptosis were analyzed by using flow cytometry assay (Becton Dickinson) after 48 h. Briefly, cells were washed twice in PBS and re-suspended in 100 μl binding buffer. Then, these cells were stained with 5 μl Annexin V-FITC and 10 μl PI (Santa Cruz Biotechnology, CA) for 10 min in a dark place at room temperature. Flow cytometry analysis was performed by a FACS can (Beckman Coulter, Fullerton, CA, USA). The data were analyzed by using FlowJo software (Treestar, Ashland, OR, USA). More information were clarified in Supplementary files.
9
Flow Cytometry-based Cell Proliferation and Apoptosis Assay
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For the flow cytometry assay for assessing cell proliferation and apoptosis, HepG2 cells were grown in 6-well plates maintaining a cell density of approximately 1×106 cells/well at standard humidified cell-culture conditions (37 °C and 5% CO2) for one day. The cultured cells were treated with the CPT loaded DNA-NWs (PI-tagged) for 48 h. The treatment group of cells were compared with the control group of cells receiving blank DNA-NWs (PI-tagged) without prior CPT loading. To make the single cell suspension, the adhered cells were detached by the trypsin treatment for 2 min at 37 ℃. The pellets of the cells obtained through the centrifugation were washed with PBS twice. The washed cells were then re-suspended in the PBS. The sample was then subjected to a flow cytometry assay (Becton Dickinson, San-Jose CA, USA) for cell counting measurements using parameters set according to the PI dye fluorescence range [4 (link)].
10
Annexin V-FITC Cell Apoptosis Assay
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The H1975 and A431 cells (1 to 5 × 105 cells/well) incubated in 6-well plates were treated by different concentrations of inhibitors for 48 h. Then, they were collected and fixed with 70% ethanol at 4 °C overnight. After beening fixed with 75% ethanol at 4 °C for 24 h, the cells were stained with Annexin V-FITC (5 μL)/propidium iodide (5 μL) and analyzed by flow cytometry assay (Becton-Dickinson, Franklin Lakes, NJ, USA).
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