The cells were stained with propidium iodide (PI) to conduct cell cycle analysis using a cell cycle staining kit (Beyotime Biotechnology, China) according to the manufacturer’s protocol and counted using a FACSCalibur flow cytometer.
Cell cycle staining kit
Manufactured by Beyotime
Sourced in China
The Cell cycle staining kit is a laboratory product designed to facilitate the analysis of the cell cycle stages. It contains reagents necessary for staining cellular DNA, enabling the identification and quantification of cells in different phases of the cell cycle, such as G0/G1, S, and G2/M.
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Lab products found in correlation
Fc500, by Beckman Coulter (2 mentions)
Sybr green rt pcr reaction kit, by Toyobo (2 mentions)
Human tnf α, by Thermo Fisher Scientific (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Facscalibur flow cytometer, by Beyotime (1 mentions)
Annexin 5 fitc pi apoptosis kit, by Beyotime (1 mentions)
Fc500, by FlowJo (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Fitc anti human cd4, by BioLegend (1 mentions)
Apc anti human cd25, by BioLegend (1 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Beyotime (1 mentions)
Cxp software, by Beckman Coulter (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Trizol plus rna purification kit, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modi ed eagle medium, by Thermo Fisher Scientific (1 mentions)
10 protocols using cell cycle staining kit
1
Cell Cycle Analysis by PI Staining
2
Cell Cycle and Apoptosis Analysis
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The cell cycle analysis was performed using a Cell Cycle Staining Kit (Beyotime Biotechnology, Shanghai, China), and cell apoptosis was assessed using an Annexin V/propidium iodide (PI) apoptosis kit (Beyotime Biotechnology). The cells were starved in RPMI 1640 without FBS for 24 h for synchronization and then seeded at a density of 105 cells/mL in six-well plates with complete medium. After 48 h, the cell cycle and apoptosis were determined by flow cytometry according to the manufacturer’s instructions.
3
Cell Cycle and Apoptosis Analysis
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Cell cycle analysis was performed using a cell cycle staining kit (Beyotime, Cat# C1052) following the manufacturer’s instructions. Cells were fixed with 70% cold ethanol for 3 h, rinsed using PBS, and then labeled with propidium iodide in the presence of RNase A for 30 min in the dark. Flow cytometry (Beckman Coulter FC500, USA) and FlowJo software (version 7.6.1) were used to determine the percentages of cells in G0/G1, S, and G2/M phases. Apoptosis was analyzed using an Annexin V-FITC/PI apoptosis kit (Beyotime, Cat# C1062M) according to the manufacturer’s instruction. Treated cells were harvested and rinsed twice with cold PBS before staining with 5 μL of Annexin V‐FITC and 5 μL of propidium iodide (PI). They were then incubated in the dark for 15 min at room temperature before analysis by flow cytometry (Beckman Coulter FC500, USA). The PHAGF apoptotic rate was determined by the proportion of Annexin V-FITC positive cells (early apoptosis) plus Annexin V-FITC/PI positive cells (late apoptosis). Cells negative for Annexin V-FITC and PI were considered live cells. Assays were done in triplicate.
4
Cell Cycle Analysis of Glioblastoma Cells
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The effect of TPL on the cell cycle in glioblastoma was analyzed using the Cell Cycle Staining Kit (Beyotime). And the cell cycle assay was performed as described previously (21 (link)). After treatment with TPL for 24h, U251 cells were trypsinized, fixed in 70% ice-cold ethanol for 24 h and washed with PBS, then incubated with 100 mg/mL RNase and 4 mg/mL propidium iodide (PI) in 500 μl PBS. The cell cycle phase was analyzed using a cytoflex (Beckman Coulter).
5
Cell Cycle and Phenotypic Analysis of NSCLC and PBMCs
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Single-cell suspensions of NSCLC cells or PBMCs were stained with the cell cycle staining kit (Beyotime), FITC anti-human CD4, APC anti-human CD25, or PE anti-human Foxp3 antibodies (Biolegend). The stained cells were washed and incubated with 1 ml intracellular fixation buffer at room temperature for 60 min. Permeabilization buffer (2 ml) were then added into each tube, and the cells were centrifuged at 1,500 rpm for 7 min. The supernatants were removed, and single-cell suspensions were analyzed by flow cytometry (Beckman, China).
6
Cell Apoptosis and Cell Cycle Analysis
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Huh7 and HepG2 cells were transfected with desired siRNA or control, then cell apoptosis assay were performed with the Annexin V-FITC/PI Apoptosis Detection Kit from the Beyotime Biotechnology (Shanghai, China). The cells were analyzed by flow cytometry (FACSCalibur flow cytometer; BD Biosciences, Franklin Lakes, NJ, USA).
The cells were stained with PI to conduct cell cycle analysis using cell cycle staining kit (Beyotime Biotechnology, China) according to the manufacturer’s protocol.
The cells were stained with PI to conduct cell cycle analysis using cell cycle staining kit (Beyotime Biotechnology, China) according to the manufacturer’s protocol.
7
Cell cycle analysis of miR-198 in RCC
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The effect of miR-198 on the cell cycle in RCC was assayed using the Cell Cycle Staining Kit (Beyotime). After transfection with miR-198 or its scrambled mimic for 48 h, A498 cells were washed with phosphate-buffered saline (PBS), trypsinized, fixed in 70% ice-cold ethanol, and incubated with 100 mg/mL RNase and 4 mg/mL propidium iodide (PI) in PBS. The cell cycle phase of the cells was analyzed using a flow cytometer (FC500; Beckman Coulter) and was analyzed using CXP software (Beckman Coulter).
8
Cell Cycle Analysis of A549 and A549/DDP Cells
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A549 and A549/DDP cells (2 × 105 cells/well) were seeded in 6‐well plates. For cell cycle distribution analysis, the cells were collected using centrifugation at 1500 rpm for 5 minutes, and the cell precipitate was washed with 70% alcohol and stored overnight in 70% alcohol. The next day, the cell precipitation was centrifuged at 1500 rpm for 5 minutes, and the supernatant was removed. Cells were stained using a Cell Cycle Staining Kit (Beyotime, China) for 30 minutes in the dark and then analysed using flow cytometry.
9
Benzyl Cinnamate Modulates Cellular Responses
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Foetal bovine serum (FBS), phosphate buffered saline (PBS), penicillin-streptomycin, trypsin-EDTA and Dulbecco’s modified Eagle medium (DMEM) were purchased from GIBCO (Grand Island, NY, USA). The human TNF-α (tumour necrosis factor-α) was purchased from Pepro Tech (Rocky Hill, NJ, USA). Griess reagent, dimethyl sulfoxide (DMSO), Cell Counting Kit-8 (CCK-8 kit), BCA Protein Assay Kit, and Annexin V-FITC/PI apoptosis kits were purchased from the BOSTER Biol.Tech. Co. (Wuhan, China). Cell Cycle Staining kits were purchased from Beyotime Biotechnology Company (Haimen, China). TRIzol™ Plus RNA Purification Kit (Invitrogen life technology, Carlsbad, CA, USA), ReverTra Ace® qPCR RT Master Mix (Beans Biol Tech. Co., Tokyo, Japan) and SYBR Green RT-PCR reaction kit (QPK-201, Toyobo, Tokyo, Japan) were used in RT-PCR experiment. Benzyl cinnamate was purchased from the Desite Biology Co., Ltd., (Chengdu, China).
10
Cytotoxic and Apoptotic Effect Analysis
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Fetal bovine serum (FBS), phosphate buffered saline (PBS), Penicillin-Streptomycin, trypsin-EDTA and Dulbecco's modi ed Eagle medium (DMEM) were purchased from GIBCO (Grand Island, NY, USA). The human TNF-α was purchased from Pepro Tech (Rocky Hill, NJ, USA). Griess reagent, dimethyl sulfoxide (DMSO), Cell Counting Kit-8 (CCK-8 kit), BCA Protein Assay Kit, and Annexin V-FITC/PI apoptosis kits were purchased from the BOSTER Biol.Tech. Co. (Wuhan, China). Cell Cycle Staining kits were purchased from Beyotime Biotechnology Company (Haimen, China). TRIzol™ Plus RNA Puri cation Kit (Invitrogen life technology, Carlsbad, CA, USA), ReverTra Ace® qPCR RT Master Mix (Beans Biol Tech. Co., Tokyo, Japan) and SYBR Green RT-PCR reaction kit (QPK-201, Toyobo, Tokyo, Japan) were used in RT-PCR experiment.
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