Facs cytometry

Manufactured by BD

Sourced in United States

FACS cytometry is a laboratory technique used to analyze and sort cells based on their physical and biochemical properties. It utilizes the principles of light scattering, wavelength emission, and fluorescence intensity to detect and characterize individual cells within a heterogeneous population.

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Lab products found in correlation

Rnase a, by Keygen Biotech (2 mentions) Annexin v 7 aad apoptosis detection kit, by Keygen Biotech (2 mentions) Fitc annexin 5 apoptosis detection kit, by BD (2 mentions) Annexin 5 propidium iodide detection kit, by Keygen Biotech (2 mentions) Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions) Cell cycle detection kit, by BD (1 mentions) Annexin 5 pe and 7 aad, by BD (1 mentions) Flow cytometry assay, by BD (1 mentions) Annexin 5 fitc kit, by BD (1 mentions) Annexin v 7 aad apoptosis detection kit, by BD (1 mentions) Annexin 5 fluorescein isothiocyanate fitc propidium iodide dual staining kit, by BD (1 mentions) Annexin 5 fitc cell apoptosis kit, by Keygen Biotech (1 mentions) Facscan, by BD (1 mentions) Propidium iodide, by Cayman Chemical (1 mentions) Alexa fluor 488 annexin 5 dead cell apoptosis kit, by Thermo Fisher Scientific (1 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) Annexin 5 fluorescein, by BD (1 mentions) Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit, by Keygen Biotech (1 mentions)

22 protocols using facs cytometry

Cell Cycle and Apoptosis Assays

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For cell cycle assay, 5 × 10⁶ cells in each group were collected and mixed with 70% cold ethanol overnight at 4°C. Cells were then washed with PBS 3 times and then incubated with PBS containing 0.5 mg/ml RNase A and 10 mg/ml propidium iodide for 30 min at 37°C in the dark. The DNA contents distribution was measured with FACS cytometry (BD Biosciences, USA). Each experiment was performed in triplicate.
For cell apoptosis assay, 5 × 10⁶ cells in each group were collected and stained with Annexin V-FITC/PI Apoptosis Detection Kit (BD Biosciences). The apoptosis rates of cells were acquired using FACS cytometry (BD Biosciences, USA). Each experiment was performed in triplicate.

Xue D., Zhou C., Shi Y., Lu H., Xu R, & He X. (2016). Nuclear transcription factor Nrf2 suppresses prostate cancer cells growth and migration through upregulating ferroportin. Oncotarget, 7(48), 78804-78812.

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Cell Cycle and Apoptosis Analysis

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After treating NSCLC cells with KPT-185 for 48 h, they were stained with propidium iodide (PI) staining buffer (Cayman Chemical, Ann Arbor, MI, USA) for 30 min at room temperature and then measured by FACS cytometry (BD Biosciences, NJ, USA). The DNA histograms were analyzed using ModFit LT cell cycle analysis software (Verify Software House).
Cell apoptosis was detected with an Alexa Fluor 488 annexin V/Dead Cell Apoptosis Kit (Invitrogen) according to the kit instructions. Briefly, after treating with KPT-185 for 48 h, the cells from both suspension and adherence were collected and co-incubated with Annexin V-fluorescein isothiocynate (FITC) and PI, then measured by FACS cytometry (BD Biosciences). The percentage of Annexin V and PI negative cells was determined based on the dot plots of FITC and PI.

Wang S., Han X., Wang J., Yao J, & Shi Y. (2014). Antitumor Effects of a Novel Chromosome Region Maintenance 1 (CRM1) Inhibitor on Non-Small Cell Lung Cancer Cells In Vitro and in Mouse Tumor Xenografts. PLoS ONE, 9(3), e89848.

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Apoptosis Analysis of Icotinib-Resistant LUAD Cells

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Icotinib-resistant LUAD cells were treated with icotinib (5 μM or 10 μM) for 72 h. Then both floating and attached cells were harvested, and staining with Annexin V-FITC Apoptosis Detection kit. Cells apoptosis were analyzed by FACS cytometry (BD Biosciences Inc., Franklin, NJ, USA).

Wu J., Zheng C., Wang Y., Yang Z., Li C., Fang W., Jin Y., Hou K., Cheng Y., Qi J., Qu X., Liu Y., Che X, & Hu X. (2021). LncRNA APCDD1L-AS1 induces icotinib resistance by inhibition of EGFR autophagic degradation via the miR-1322/miR-1972/miR-324-3p-SIRT5 axis in lung adenocarcinoma. Biomarker Research, 9, 9.

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Cell Cycle and Apoptosis Analysis Protocol

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For cell cycle analysis, cells were seeded into 6-well plates, and then trypsinized and centrifuged. After washing in PBS, cells were treated with 70% of ice-cold ethanol for 2 h all night. The cell cycle detection kit (BD Bioscience, San Jose, CA, USA) was added for 30 min. For cell apoptosis analysis, cells in cold PBS were incubated with 100 μL of 1× binding buffer and 5 μL of Annexin V-PE and 7AAD (BD Biosciences) for 15 min in the dark. Both were analyzed by FACS cytometry (BD Biosciences).

Huang J., Pan B., Xia G., Zhu J., Li C, & Feng J. (2020). LncRNA SNHG15 regulates EGFR-TKI acquired resistance in lung adenocarcinoma through sponging miR-451 to upregulate MDR-1. Cell Death & Disease, 11(7), 525.

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Cell Cycle and Apoptosis Analysis

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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.

Zhu L., Dai L., Yang N., Liu M., Ma S., Li C., Shen J., Lin T., Wang D., Pan W, & Li X. (2020). Transcription factorIRX5 promotes hepatocellular carcinoma proliferation and inhibits apoptosis by regulating the p53 signalling pathway. Cell Biochemistry and Function, 38(5), 621-629.

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Cell Apoptosis Analysis with Annexin V

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The different treatment of T98G, LN229, T98G/TR and LN229/TR cells was collected for staining. According to the standard protocols, the cell apoptosis was determined by an Annexin‐V‐FITC Kit (BD Pharmingen, San Diego, CA, USA). After washing with PBS, Annexin V/propidium iodide was added to cells and incubated for 30 min in the dark, and the apoptotic rate was measured by FACS cytometry (BD Biosciences, USA).

Chen Y., Wang Y., Qiu K., Cao Y., Zhang F., Zhao H, & Liu X. (2022). YTHDF2 promotes temozolomide resistance in glioblastoma by activation of the Akt and NF‐κB signalling pathways via inhibiting EPHB3 and TNFAIP3. Clinical & Translational Immunology, 11(5), e1393.

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5-FU-Induced Apoptosis Assay

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The apoptosis assay was done with the AnnexinV-7AAD apoptosis detection kit (KeyGEN BioTECH, Nanjing, Jiangsu Province, China) according to the manufacturer’s instructions. To detect the effect of PTTG3P over-expression on 5-FU-induced cell apoptosis, Lv-PTTG3P and Lv-con cells were seeded in 6-well plates and treated on the following day with 5-FU. After incubation for 48 h, cells were harvested, stained using AnnexinV-7AAD apoptosis detection kit and then analyzed by FACS cytometry (BD Biosciences, San Jose, CA, USA).All experiments were performed in duplicate and reproducibility was checked in three independent experiments.

Huang J.L., Cao S.W., Ou Q.S., Yang B., Zheng S.H., Tang J., Chen J., Hu Y.W., Zheng L, & Wang Q. (2018). The long non-coding RNA PTTG3P promotes cell growth and metastasis via up-regulating PTTG1 and activating PI3K/AKT signaling in hepatocellular carcinoma. Molecular Cancer, 17, 93.

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Annexin-V/PI Apoptosis Assay

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On the basis of protocol, cell apoptosis was monitored with Annexin-V fluorescein isothiocyanate (FITC)/propidium iodide (PI) dual-staining kit (BD Biosciences, San Jose, CA). In brief, after treating with FITC-Annexin-V and PI in succession for 15 min in the dark, the apoptotic cells were analyzed using FACS cytometry (BD Biosciences).

Zhou T., Wu L., Ma N., Tang F., Yu Z., Jiang Z., Li Y., Zong Z, & Hu K. (2020). SOX9-activated FARSA-AS1 predetermines cell growth, stemness, and metastasis in colorectal cancer through upregulating FARSA and SOX9. Cell Death & Disease, 11(12), 1071.

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Cell Cycle and Apoptosis Analysis

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After treating NSCLC cells with chidamide or icotinib alone, or in combination for 48h, cells were washed with phosphate buffer saline (PBS) solution, and incubated in DNA staining solution and permeabilization solution for 30min at room temperature according to the kit instructions (MultiSciences (LIANKE) Biotech Co., Ltd) and then measured by FACS cytometry (BD Biosciences, NJ, USA). The DNA histograms were analyzed using ModFit LT cell cycle analysis software (Verify Software House).
Cell apoptosis was performed using Annexin V-FITC cell apoptosis kit (Keygen Biotech, Nanjing, China) according to the kit instructions. Briefly, after treating with chidamide or icotinib alone, or in combination for 48h, the cells from both suspension and adherence were collected and co-incubated with Annexin V-fluorescein isothiocyanate (FITC) and PI, then measured on a Becton-Dickinson FACScan instrument (BD Biosciences, NJ, USA) using Cell Quest software. This analysis allows discrimination between viable (annexin V-/PI-), early (annexin V+/PI-) and late (annexin V+/PI+) apoptotic cells, as well as necrotic and/or part of late apoptotic (annexin V-/PI+) cells.

Zhang N., Liang C., Song W., Tao D., Yao J., Wang S., Ma L., Shi Y, & Han X. (2019). Antitumor activity of histone deacetylase inhibitor chidamide alone or in combination with epidermal growth factor receptor tyrosine kinase inhibitor icotinib in NSCLC. Journal of Cancer, 10(5), 1275-1287.

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Apoptosis Assay of KPT-185 in Cancer Cells

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PC9 and PC9GR cells in logarithmic growth stage were seeded in six-well plates at a density of 2 × 10⁵ cells per well and grown overnight. The next day, the growth medium was replaced with fresh media with multiple dilution concentrations of KPT-185 at 37°C for 48 h. The cells from both suspension and adherence were collected and resuspended in binding buffer containing Annexin V-fluorescein isothiocynate (FITC). Staining solution with propidium iodide (PI) was then added following the kit instructions, and localization of Annexin V and PI for apoptotic cells was performed by FACS cytometry (BD Biosciences, Franklin Lakes, NJ, USA) and percentage of apoptotic cells were obtained.

Wei N., Song Y., Zhang F., Sun Z, & Zhang X. (2020). Transcriptome Profiling of Acquired Gefitinib Resistant Lung Cancer Cells Reveals Dramatically Changed Transcription Programs and New Treatment Targets. Frontiers in Oncology, 10, 1424.

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