Facsort flow cytometer

Manufactured by Beckman Coulter

Sourced in United States

The FACSort Flow Cytometer is a high-performance instrument designed for cell analysis and sorting. It utilizes flow cytometry technology to rapidly detect, identify, and sort different cell types based on their physical and biochemical characteristics. The FACSort provides precise quantification and detailed analysis of multiple parameters for each individual cell within a sample.

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Lab products found in correlation

Annexin 5 fitc apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Anti cd206 clone 19, by BD (1 mentions) Propidium iodide staining solution, by Vazyme (1 mentions) Annexin 5, by Vazyme (1 mentions)

10 protocols using facsort flow cytometer

Apoptosis Assay for Human Colon Cancer

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After treatment, human colon cancer cell line with HCT-116 WT, p53^−/− (HCT-116 p53-KO), DLD1 and Bax^−/− (HCT116 Bax-KO) were suspended in 1 × 10⁵ cells/ml, and 5 μl Annexin V and 5 μl propidium iodide staining solution were added to 100 μl of the cell suspension. Then added 400 μl Binding Buffer to the cell suspension again. After the cells were incubated at room temperature for 10 min in the dark, stained cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, USA). Cell debris was excluded from the analysis by an appropriate forward light scatter threshold setting. Compensation was used wherever necessary.

Liu Y., Huang Y., Ding J., Liu N., Peng S., Wang J., Wang F, & Zhang Y. (2019). Targeting Akt by SC66 triggers GSK-3β mediated apoptosis in colon cancer therapy. Cancer Cell International, 19, 124.

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Flow Cytometric Apoptosis Quantification

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Cells were suspended at 1 × 10⁶ cells/ml, and 5 μl of Annexin V and propidium iodide staining solution were added to 200 μl of the cell suspension. After the cells were incubated at room temperature for 15 min in the dark, stained cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, U.S.A.). Cell debris was excluded from the analysis using an appropriate forward light scatter threshold setting. Compensation was used whenever necessary.

Zhu T., Chen J., Zhao Y., Zhang J., Peng Q., Huang J., Luo J, & Zhang W. (2019). Neuromedin B mediates IL-6 and COX-2 expression through NF-κB/P65 and AP-1/C-JUN activation in human primary myometrial cells. Bioscience Reports, 39(10), BSR20192139.

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Apoptosis Analysis of Colon Cancer Cells

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Human colon cancer cell line with HCT116 WT, p53^−/− (HCT116 p53-KO), PUMA^−/− (HCT116 PUMA-KO), and Bax^−/− (HCT116 Bax-KO) were suspended in 1 × 10⁵ cells/ml, and 5 μl Annexin V and 5 μl propidium iodide staining solution were added to 100 μl of the cell suspension. Then 400 μl binding buffer was added to cell suspension again. After the cells were incubated at room temperature for 10 min in the dark, stained cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, USA). Cell debris was excluded from the analysis by an appropriate forward light scatter threshold setting. Compensation was used wherever necessary.

Sun L., Huang Y., Liu Y., Zhao Y., He X., Zhang L., Wang F, & Zhang Y. (2018). Ipatasertib, a novel Akt inhibitor, induces transcription factor FoxO3a and NF-κB directly regulates PUMA-dependent apoptosis. Cell Death & Disease, 9(9), 911.

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Apoptosis Quantification in Colon Cancer Cells

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HCT‐116 and LoVo cells were suspended in 1 × 10⁶ cells/mL, and 5 μL of Annexin V and propidium iodide staining solution were added to 300 μL of the cell suspension. After the cells were incubated at room temperature for 15 min in the dark, stained cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, USA). Cell debris was excluded from the analysis by an appropriate forward light scatter threshold setting. Compensation was used wherever necessary.

Chen J., Zhong J., Liu Y., Huang Y., Luo F., Zhou Y., Pan X., Cao S., Zhang L., Zhang Y, & Wang J. (2018). Purified vitexin compound 1, a new neolignan isolated compound, promotes PUMA‐dependent apoptosis in colorectal cancer. Cancer Medicine, 7(12), 6158-6169.

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Apoptosis Assay of Colon Cancer Cells

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For flow cytometry analysis (FACS analysis), human colon cancer cell lines with HCT116, RKO, SW48 were harvested in 1 × 10⁵ cells/mL after using different treatment. These groups were suspended by 100 μL binding buffer, and 5 μL Annexin V and 5 μL propidium iodide staining solution were added to the cell suspension respectively. Later, added 400 μL binding buffer into the cell suspension again. At room temperature, the cells were incubated in the dark for 10 min, and then cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, USA) at 488 nm. Fluorescent emission of FITC was measured at 515–545 nm and that of DNA-PI complexes at 564–606 nm. Cell debris was excluded from the analysis by an appropriate forward light scatter threshold setting. Compensation was used wherever necessary.

Liu J., Long S., Wang H., Liu N., Zhang C., Zhang L, & Zhang Y. (2019). Blocking AMPK/ULK1-dependent autophagy promoted apoptosis and suppressed colon cancer growth. Cancer Cell International, 19, 336.

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Annexin V-FITC/PI Apoptosis Assay

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Cell apoptosis assay by Annexin V-FITC (fluorescein isothiocyanate)/propidium iodide (PI) staining The CMs apoptosis was measured using Annexin V-FITC Apoptosis Detection Kit (Invitrogen Corporation), as our established method21 . Briefly, cultured cells (at a density of 3 × 10⁵ /ml) were washed twice with PBS and incubated in 500 μl binding buffer containing 5 μl of Annexin V-FITC and 5 μl of PI in the dark for 15 min at room temperature. The stained samples were then analyzed on a FACSort flow cytometer within an hour following the manufacturer’s protocol (Beckman Coulter, Fullerton, USA).

Wang L., Shen M., Guo X., Wang B., Xia Y., Wang N., Zhang Q., Jia L, & Wang X. (2017). Volume-sensitive outwardly rectifying chloride channel blockers protect against high glucose-induced apoptosis of cardiomyocytes via autophagy activation. Scientific Reports, 7, 44265.

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Peritoneal Macrophage Phenotyping by Flow Cytometry

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To identify and measure the proportion of peritoneal cell types, markers on peritoneal macrophages from patients with CAPD were measured by triple colour flow cytometry using a FACSort flow cytometer (Beckman Coulter, Inc.). Anti-CD14 (clone M5E2; cat. no. 550787; 1:100; BD Biosciences), anti-CCr7 (clone 3D12; cat. no. 552176; 1:50; BD Biosciences) and anti-CD206 (clone 19.2; cat. no. 551135; 1:50; BD Biosciences) antibodies were labelled with PerCP, PE, and FITC, respectively. Class-matched isotype immunoglobulin PerCP, PE and FITC-conjugated negative control monoclonal antibodies were added to individual tubes for all samples to identify nonspecific binding. The broad macrophage gate was refined by plotting side scatter against CD14. In patients with CAPD, CD14- and CCr7-positive cells were considered M1 macrophages, and CD14- and CD206-positive cells were considered M2 macrophages. Murine macrophages (J774A.1 cells) cultured in different concentrations of glucose were considered M1 or M2 cells based on the expression of CCr7 or CD206, respectively. The flow cytometry data were analysing using WinMDI software (version 2.9; The Scripps Research Institute).

Lin J., Kong Q., Hao W, & Hu W. (2020). High glucose contributes to the polarization of peritoneal macrophages to the M2 phenotype in vivo and in vitro. Molecular Medicine Reports, 22(1), 127-134.

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Annexin V-PI Apoptosis Assay

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Cells were suspended at 1×10⁶ cells/mL, and 5 µL of annexin V and propidium iodide staining solution was added to 200 µL of the cell suspension. After the cells were incubated at room temperature for 15 min in the dark, cells were centrifuged at 1000 g for 5 min and resuspended in 200 µL phosphate-buffered saline. Stained cells were then assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, California, USA).

Chen J., Shi Y., Huang J., Luo J, & Zhang W. (2020). Neuromedin B receptor mediates neuromedin B-induced COX-2 and IL-6 expression in human primary myometrial cells. Journal of Investigative Medicine, 68(6), 1171-1178.

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Annexin V and Propidium Iodide Cell Apoptosis Assay

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Human colon cancer cell lines HCT116 WT and HCT116 p53–/– were suspended in 1 × 10⁵ cells/ml, and 5 μl Annexin V and 5 μl propidium iodide staining solution were added to 100 μl of the cell suspension. Then 400 μl binding buffer was added to cell suspension again. After the cells were incubated at room temperature for 10 min in the dark, stained cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, USA). Cell debris was excluded from the analysis by an appropriate forward light scatter threshold setting. Compensation was used wherever necessary.

Zhang Y., Zhang C., Li J., Jiang M., Guo S., Yang G., Zhang L., Wang F., Yi S., Wang J., Fu Y, & Zhang Y. (2022). Inhibition of AKT induces p53/SIRT6/PARP1-dependent parthanatos to suppress tumor growth. Cell Communication and Signaling : CCS, 20, 93.

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Annexin V and PI Apoptosis Assay

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The B16F10 cells treated with PBS, Dox (1 μM) and Dox-coated hydrogel with or without R837 (10 μg/mL) were suspended in 1 × 10⁵ cells/mL, and 5 μL Annexin V (Vazyme, Nanjing, China) and 5 μL propidium iodide staining solution (Vazyme, Nanjing, China) were added to 100 μL of the cell suspension. 400 μL binding buffer was next added to the cell suspension. After incubated at room temperature for 10min in the dark, the stained cells were assayed and quantified using a FACSort Flow Cytometer (Beckman Coulter, Brea, CA, USA). Cell debris was excluded from the analysis by an appropriate forward light scatter threshold setting. What's more, compensation was used wherever necessary.

Li J., Luo G., Zhang C., Long S., Guo L., Yang G., Wang F., Zhang L., Shi L., Fu Y, & Zhang Y. (2022). In situ injectable hydrogel-loaded drugs induce anti-tumor immune responses in melanoma immunochemotherapy. Materials Today Bio, 14, 100238.

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