The human chorionic trophoblast cell line HTR8/SVneo was obtained from the American Type Culture Collection (Manassas, VA, USA) and preserved in our laboratory. Reagents and equipment included fetal bovine serum (FBS; Gibco, USA), dimethyl sulfoxide (Sigma, USA), RPMI 1640 medium (HyClone, USA), RPMI 1640 medium, no glucose (Thermo, Germany), pancreatin (Gibco, USA), penicillin–streptomycin solution (HyClone, USA), d -mannitol (Solarbio, USA), Cell Cycle Detection Kit (Shanghai Bestbio Company), phosphate-buffered saline (PBS; HyClone, USA), glucose powder (Hefei Bomei Biotechnology Co., Ltd.), Cell Cycle Detection Kit (Sigma Company, USA), Cell Apoptosis Detection Kit (Thermo, Germany), Cell Counting Kit-8 (CCK-8; Beyotime Biotechnology, Shanghai, China), absolute ethanol (Sinopharm Group Co., Ltd., Beijing, China), adjustable pipette (2.5–1,000 μL), desktop refrigerated ultracentrifuge (5424R), high-speed mini centrifuge (Eppendorf, Germany), Vortex oscillator (Vortex-Genie®, Science Industries, Inc.), CO2 incubator, −80°C refrigerator, thermostatic water bath (Thermo, Germany), electronic platform scale (Sartorius, Germany), laminar flow bench (Suzhou Antai Airtech Co., Ltd.), inverted microscope (Olympus, Japan), microplate reader (Promega, USA), pure water system (Millipore, USA), and flow cytometer (BD FACSAria™ III (BD FACS AriaIII/BD, USA).
Cell cycle detection kit
Manufactured by Merck Group
Sourced in United States
The Cell Cycle Detection Kit is a laboratory tool designed to analyze the distribution of cells within the different phases of the cell cycle. It provides a standardized method for quantifying the relative proportions of cells in the G0/G1, S, and G2/M phases based on their DNA content. The kit includes reagents and protocols to prepare and stain cell samples for flow cytometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Co2 incubator, by Thermo Fisher Scientific (3 mentions)
Facs cytometry, by Merck Group (2 mentions)
Pure water system, by Merck Group (1 mentions)
D mannitol, by Solarbio (1 mentions)
Pancreatin, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Facsaria 3, by BD (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Inverted microscope, by Olympus (1 mentions)
Absolute ethanol, by Sinopharm (1 mentions)
Htr 8 svneo, by American Type Culture Collection (1 mentions)
Microplate reader, by Promega (1 mentions)
Rnase a, by Thermo Fisher Scientific (1 mentions)
Guava easycyte ht flow cytometer, by Merck Group (1 mentions)
Modfitlt version 3, by Verity Software House (1 mentions)
Facscalibur, by BD (1 mentions)
Facscan, by BD (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Annexin 5 fluorescein isothiocyanate fitc kit, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
9 protocols using cell cycle detection kit
1
Cell Cycle and Apoptosis Analysis of HTR8/SVneo Cells
2
Cell Cycle and Apoptosis Analysis
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The H1299 and A549 cell lines were harvested with trypsin (under 37°C) and centrifugation at 300 × g (at room temperature for 5 min), and stained with either a cell cycle detection kit (containing PI; Sigma-Aldrich; Merck KGaA), with RNase A (Fermentas; Thermo Fisher Scientific, Inc.) or with Annexin V-APC for cell apoptosis detection (AAT Bioquest Inc.), according to the manufacturer's instructions. For cell cycle detection, the cells were stained at room temperature for 20 min, while for apoptosis analysis, the cells were incubated at room temperature for 60 min. Both experiments required the avoidance of light. A Guava easyCyte HT flow cytometer (EMD Millipore) was used to perform the experiments and the FlowJo software (v10.0.7; FlowJo, LLC) was used to analyze the cell cycle and apoptosis data. Each group of samples included three biological repeats for data analysis.
3
Cell Cycle Analysis of SMMC7721 Cells
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SMMC7721 cells were fixed with 70% ethanol overnight at 4°C and were collected the next day and washed twice with PBS. Then, cells were treated with 0.5 mg/ml RNase (Sigma) for 30 min and stained with propidium iodide (PI) using a Cell Cycle Detection kit (Sigma) to analyze the cell cycle at room temperature in the dark for 30 min according to the manufacturer's instructions. We counted 14,000 events for each PI-stained sample on a FACS Calibur (BD Biosciences, Franklin Lakes, NJ, USA) and analyzed the percentage of cells in the G1, S, and G2/M phases using a FACScan (BD Biosciences, San Jose, CA, USA) flow cytometer. ModFit LT version 3.2 software (Verity Software House; Topsham, Devon, UK) was used to analyze the cell cycle data.
4
Taraxerol Anti-Cancer Assay Protocol
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Taraxerol (purity > 96%) was gained from the New Drug Research and Development Center of North China Pharmaceutical Group Corporation (Hebei, China). The Roswell Park Memorial Institute (RPMI)-1640 medium and fetal bovine serum (FBS) were purchased from Gibco-BRL (Invitrogen Life Technologies, USA). 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT), dimethyl sulfoxide (DMSO), the Annexin V-fluorescein isothiocyanate (FITC) kit, and the cell cycle detection kit were purchased from Sigma (St. Louis, USA).
5
Cell Cycle and Apoptosis Analysis
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Cell cycle and apoptosis assays were performed using flow cytometry. CMM cells transfected with siRNA or vector were cultivated in six-well plates. Cells were treated with or without irradiation for 48 h, and then harvested by centrifugation. An annexin V-fluorescein isothiocyanate/propidium iodide staining kit (MB-CHEM Mumbai, India) and cell cycle detection kit (Sigma) were used to stain the cells, according to the manufacturers’ protocols. All the tests were performed in triplicates.
6
Evaluating P-gp Inhibition and Cell Viability
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PTX and FKA were purchased from Sigma (St. Louis, MO, USA). Sodium aescinate (Aes) (≥98%) was purchased from Meilunbio® (Dalian, China). Monoclonal rabbit anti-human P-gp (cat. no. 13978) was purchased from Cell Signaling Technology (MA, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies (Kumamoto, Japan). Annexin V‑fluorescein isothiocyanate/propidium iodide Apoptosis Detection Kit was purchased from BD Biosciences (Franklin Lakes, NJ, USA). The cell cycle detection kit was purchased from Sigma (St. Louis, MO, USA). Dialysis membranes (MWCO 1000) were obtained from the Shanghai Medical Chemical Reagent Co., Ltd. (Shanghai, China). Phosphate buffer and 0.25% trypsin were purchased from Gibco, Invitrogen Corp (Ontario, USA).
7
CCK-8 Assay and Cell Cycle Analysis of HepG2 Cells
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For the CCK-8 assay, HepG2 cells were seeded in 96-well plates at a density of 2,000 cells per well and incubated for 1, 2, 3, 4, or 5 days. Then, 10 μL of CCK-8 (Dojindo Molecular Technologies, Japan) was added to each well, incubated for 4 h, and mixed gently on an orbital shaker for 2 min before the absorbance value (OD) of each well was measured at 450 nm. For the cell cycle assay, the cells were seeded on 6 cm diameter plates with DMEM containing 10% FBS and labeled using a cell cycle detection kit (Sigma, USA) following the manufacturer’s protocols. The DNA content of labeled cells was analyzed with FACS cytometry (Millipore, USA). The experiments were performed in triplicate.
8
Cell Cycle and Apoptosis Analysis
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Cells were seeded on 6 cm-diameter plates with RPMI-1640 containing 10% FBS. After treatment, cells were labeled by using a cell-cycle detection Kit (Sigma, United States) and annexin V-FITC/PI staining kit (eBioscience, United States), according to manufacturer’s instructions. The DNA content of labeled cells was analyzed with FACS cytometry (Millipore, United States). Experiments were performed in triplicate.
9
Cell Cycle and Apoptosis Assays
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Cell cycle and apoptosis assays were performed using ow cytometry. CMM cells transfected with siRNA or vector were cultivated in 6-well plates. Cells were treated with or without irradiation or additional processing for 48 h, and then harvested by centrifugation. An annexin V-uorescein isothiocyanate (FITC)/propidium iodide (PI) staining kit (Mbchem) and cell cycle detection kit (Sigma) were used to stain the cells, according to the manufacturers' protocols. All the tests were performed in triplicates.
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