Facs analyzer

Manufactured by BD

Sourced in United States

The FACS analyzer is a specialized laboratory instrument used for the analysis and sorting of cells or other microscopic particles. It utilizes fluorescence-activated cell sorting (FACS) technology to precisely measure and separate different cell types based on their physical and biochemical characteristics.

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Lab products found in correlation

Fitc annexin 5 apoptosis detection kit, by BD (7 mentions) Cell cycle assay kit, by Keygen Biotech (3 mentions) Pcbascei, by Addgene (2 mentions) Pimej5gfp, by Addgene (2 mentions) Pdrgfp, by Addgene (2 mentions) Phosflow fix 1, by BD (2 mentions) Golgistop, by BD (2 mentions) Transwell plate, by Thermo Fisher Scientific (1 mentions) Annexin 5 apc pi apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Magnetic cell sorting, by Miltenyi Biotec (1 mentions) Il 17a, by BioLegend (1 mentions) Ifn γ, by BioLegend (1 mentions) Cd8 pe cy7 clone sk1, by BioLegend (1 mentions) Modfit lt, by Verity Software House (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Annexin 5 fluorescein isothiocyanate fitc, by Merck Group (1 mentions) Propidium iodide, by Merck Group (1 mentions) Dnase free rnase a, by Merck Group (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)

31 protocols using facs analyzer

Cell Apoptosis and Cycle Analysis

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Apoptosis detection assay was performed using Annexin V-FITC Apoptosis Detection Kits (BD, USA) using a FACS analyzer (BD, USA) in accordance with the manufacturer's experiment procedures. After A498 and 786O cells were collected and washed in PBS for three times, 500 ul cell suspension, 5 ul Annexin V-FITC, and 5 ul propidium iodide (PI) solution were resuspended in each collection tube. Flow cytometry was performed to measure the effect of different groups on cell cycle distribution of A498 and 786O cells. After A498 and 786O cells were collected and washed in PBS for three times, and disposed using cell cycle assay kit (Nanjing KeyGen Biotech), the percentage of the cells number of each cell cycle phase (G0/G1, S, G2M) was detected using a FACS analyzer (BD, USA).

Xu W., Liu W.R., Xu Y., Tian X., Anwaier A., Su J.Q., Zhu W.K., Shi G.H., Wei G.M., Huang Y.P., Qu Y.Y., Zhang H.L, & Ye D.W. (2021). Hexokinase 3 dysfunction promotes tumorigenesis and immune escape by upregulating monocyte/macrophage infiltration into the clear cell renal cell carcinoma microenvironment. International Journal of Biological Sciences, 17(9), 2205-2222.

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Evaluating miR-187-3p Impact on Cell Cycle

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Flow cytometry was performed to measure the effect of miR-187-3p inhibitor and mimics interference on cell cycle distribution of A498 and 786O cells in comparison with NC group. After A498 and 786O cells were collected and washed in PBS for three times, and disposed using cell cycle assay kit (Nanjing KeyGen Biotech), the percentage of the cells number of each cell cycle phase (G0/G1, S, G2M) was detected using a FACS analyzer (BD, USA).

Xu W., Liu W., Anwaier A., Tian X., Su J., Shi G., Wei S., Qu Y., Zhang H, & Ye D. (2022). Deciphering the role of miR-187-3p/LRFN1 axis in modulating progression, aerobic glycolysis and immune microenvironment of clear cell renal cell carcinoma. Discover. Oncology, 13, 59.

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Apoptosis Detection by Flow Cytometry

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Apoptosis detection assay was performed using Annexin V-FITC Apoptosis Detection Kits (BD, USA) using a FACS analyzer (BD) in accordance with the manufacturer's experiment procedures. After A-498 and 786-O cells were collected and washed in PBS for three times, 500ul cell suspension, 5ul Annexin V-FITC, and 5ul propidium iodide (PI) solution were resuspended in each collection tube. Annexin V-FITC (-) PI (-), E3, stands for normal cells; Annexin V-FITC (+) PI (-), E4, stands for early apoptotic cells; Annexin V-FITC (+) PI (+), E2, stands for late apoptotic cells; Annexin V-FITC (-) PI (+), E1, stands for mechanical necrotic cells. Both early and late apoptotic cell (E4 and E2) were counted and measured as apoptotic cells.

Liu W., Ma C., Xu H., Wang L., Xu W., Zhang H., Wang Z., Li J., Zhang J., Liu X., Zhao S, & Wang T. (2022). miR-184-5p inhibits cell proliferation, invasion and predicts prognosis of clear cell renal cell carcinoma by targeting NUS1 dehydrodolichyl diphosphate synthase subunit: Results from large-scale comprehensive identification and validation. Journal of Cancer, 13(5), 1398-1409.

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Quantification of Apoptosis via Annexin V Assay

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Apoptosis was detected using Annexin V-FITC Apoptosis Detection Kit (BD, USA) in accordance with the manufacturer’s procedures. Briefly, after transfection with miRNA mimics, the miRNA inhibitor, or the control, FaDu and HSC-3 cells were washed three times with phosphate-buffered saline (PBS; BD, USA) and then added to 500 μl of 1× binding buffer. Subsequently, 500 μl of cell suspension, 5 μl of Annexin V-FITC, and 5 μl of propidium iodide solution were added to each collection tube. After incubation for 15 min, apoptotic cells were detected using a FACS Analyzer (BD).

Li J., Chen W., Luo L., Liao L., Deng X, & Wang Y. (2020). The microRNA miR-29c-5p inhibits cell proliferation and migration by targeting TMEM98 in head and neck carcinoma. Aging (Albany NY), 13(1), 769-781.

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Apoptosis and Cell Cycle Analysis

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Apoptosis and cell-cycle distribution were assessed using flow cytometry. After incubation with Adriamycin (6 µg/mL) for 24 h, the cells were harvested with ethylenediaminetetraacetic acid (EDTA)-free trypsin and washed twice with a D-Hanks solution. Then, a 5 µL Annexin V-APC assay was added for 15 min at an ambient temperature and shielded from light. Data analysis was carried out using a fluorescence-activated cell sorting (FACS) analyzer (BD Biosciences, USA).
Following the same procedure described for the cellular apoptosis assessment, the cells were washed and a permeabilization (10 µL) and propidium iodide (PI) staining solution (5 µL) were added. Following incubation for 30 min away from light and at an ambient temperature, the cell-cycle distribution was analyzed using the FACS analyzer. The experiments were performed in triplicate.

Zhang J.T., Chen J., Ruan H.C., Li F.X., Pang S., Xu Y.J., Huang D.L, & Wu X.H. (2021). Microribonucleic Acid-15a-5p Alters Adriamycin Resistance in Breast Cancer Cells by Targeting Cell Division Cycle-Associated Protein 4. Cancer Management and Research, 13, 8425-8434.

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Homologous Recombination Efficiency Assay

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Cells were transfected using Lipofectamine LTX with: pCDNA mCherry-C1, pDRGFP (Addgene#26475) or pimEJ5GFP (Addgene#44026), and pCBA SceI (Addgene#26477), which were kind gifts from Maria Jasin or Jeremy Stark. Cells were sorted on a FACS analyzer (BD Biosciences) to determine the population of GFP positive cells normalized to mCherry-C1 expression to account for differences in transfection efficiency (Bennardo et al., 2008 (link); Pierce et al., 1999 (link)).

Sarraf S.A., Sideris D.P., Giagtzoglou N., Ni L., Kankel M.W., Sen A., Bochicchio L.E., Huang C.H., Nussenzweig S.C., Worley S.H., Morton P.D., Artavanis-Tsakonas S., Youle R.J, & Pickrell A.M. (2019). PINK1/Parkin Influences Cell Cycle by Sequestering TBK1 at Damaged Mitochondria, Inhibiting Mitosis. Cell reports, 29(1), 225-235.e5.

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Cell Cycle Analysis by Flow Cytometry

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Cell cycle analysis was performed using flow cytometry. For DNA content analysis, approximately 10⁶ cells were fixed in 80% ethanol for at least 1 h at 4°C. Ethanol-fixed cells were stained with propidium iodide (PI) staining solution (50 μg/ml PI, 0.1 mg/ml RNase A, 0.1% NP-40, 0.1% trisodium citrate) for 30 min, and analyzed using a FACS analyzer (BD Biosciences, San Diego, CA, USA).

Kim J.K., Jung H.J., Hyun M., Lee J.Y., Park J.H., Suh S.I., Baek W.K, & Kim H.A. (2023). Resistance of hypervirulent Klebsiella pneumoniae to cathepsin B-mediated pyroptosis in murine macrophages. Frontiers in Immunology, 14, 1207121.

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Glioma Cell Apoptosis via hucMSCs-sTRAIL

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In this study, the effect of hucMSCs infected with LV-scFv-sTRAIL on U87G apoptosis was investigated using co-culture assay. For co-culture, U87G cells (1 × 10⁵/well) were plated in the lower chamber of the transwell plate (Thermo Fisher Scientific) and incubated for 24 h. Then, hucMSCs (1 × 10⁴/well) in each group were grown in the upper chamber. After another 48 h, glioma cell apoptosis was detected using the Annexin V-APC/PI apoptosis detection kit (Invitrogen, Carlsbad, CA, United States). Briefly, the cells were resuspended in 300 µl binding buffer, followed by the addition of Annexin V-APC solution (5 µl). After 25 min of incubation at 4°C, the cells were resuspended for 10 min in a binding buffer with 5 μl of PI. Apoptotic cells were counted using a FACS analyzer (BD Biosciences).

Xue T., Wang X., Ru J., Zhang L, & Yin H. (2022). The inhibitory effect of human umbilical cord mesenchymal stem cells expressing anti-HAAH scFv-sTRAIL fusion protein on glioma. Frontiers in Bioengineering and Biotechnology, 10, 997799.

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B Cell-Mediated Immunomodulation in EAMG

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Splenic CD1d^hiCD5⁺ B cells from donor mice in the PBS/EAMG (AChR-immunized mice receiving PBS) and in the GM-CSF/EAMG (AChR immunized mice receiving GM-CSF) groups were isolated and co-cultured with responder T or B cells from immunized mice at 1:1 ratio. For proliferation assay, responder CD4⁺ or CD19⁺cells were isolated from mice in untreated EAMG mice using magnetic cell sorting (Miltenyi Biotec, Auburn, CA) and were stained with CFSE at a concentration of 1 µM for 10 min at 37 ^oC. Cells were washed three times and plated into 96-well, flat-bottom plates at 5 × 10⁵ cells/well. T cell-depleted enriched DCs (1 × 10⁵ cells/well) (also accomplished by magnetic cell sorting) were used as feeder cells in these studies. Cells were stimulated with tAChR (5 µg/ml) for 72 hrs, and then harvested for CFSE dilution studies and intracellular cytokine expression using a FACS analyzer (BD Biosciences). For antibody production, total anti-AChR IgG concentrations in culture supernatants in B cell co-cultures were measured using a mouse IgG enzyme-linked immunosorbent assay (ELISA) set (Bethyl Laboratories, Montgomery, TX), according to the manufacturer’s specifications.

Sheng J.R., Quan S, & Soliven B. (2014). CD1dhiCD5+ B cells Expanded by GM-CSF in Vivo Suppress Experimental Autoimmune Myasthenia Gravis. Journal of immunology (Baltimore, Md. : 1950), 193(6), 2669-2677.

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Cell Cycle Analysis of miR-184-5p Modulation

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Flow cytometry was performed to measure the effect of miR-184-5p inhibitor and mimics interference on cell cycle distribution of A-498 and 786-O cells in comparison with NC group. After A-498 and 786-O cells were collected and washed in PBS for three times and disposed using cell cycle assay kit (KeyGen Biotech, Nanjing, China), the percentage of the cells number of each cell cycle phase (G0/G1, S, G2M) was detected using a FACS analyzer (BD, USA).

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