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Facs arial

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The FACS Arial is a flow cytometry instrument designed for cell analysis and sorting. It is capable of detecting and analyzing multiple parameters of individual cells or particles in a sample. The FACS Arial provides accurate and reliable data for researchers working in fields such as immunology, cell biology, and cancer research.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Arc antibody, by Bioss Antibodies (2 mentions) Smrter stranded total rna seq kit v2 pico input mammalian components kit, by Takara Bio (2 mentions) Neun antibody, by Abcam (2 mentions) Propidium iodide, by Keygen Biotech (1 mentions) Cd11b, by Abcam (1 mentions)

6 protocols using facs arial

1

Cell Cycle and Apoptosis Analysis

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Flow cytometry was used to assess the cell cycle and apoptosis. For the cell cycle assay, SW-480 and DLD-1 cells (1×106 cells/well) were seeded in six-well plates in DMEM with 10% FBS and cultured for 48 h. Following trypsinization and washing with PBS, cells were fixed in 70% ethanol at 4°C for 2 h. Subsequently, the samples were resuspended in the staining buffer with propidium iodide (PI; Nanjing KeyGen Biotech Co., Ltd.) and RNase A (the ratio of PI to RNase A was 9:1), and incubated in the dark for 30 min at room temperature. Finally, the samples were analyzed using a flow cytometer (FACS Arial, BD Biosciences). The proportion of cells in the S + G2 phase was calculated using FlowJo v10.8.1 software (BD Biosciences).
For the apoptosis assay, the cells were harvested and incubated with Annexin V-APC and PI using an Annexin V-APC/PI Apoptosis Detection kit (Bio-Rad Laboratories, Inc.), according to the manufacturer's instructions, and analyzed using a flow cytometer (as described above).
Liu P., Liu J., Ding M., Liu Y., Zhang Y., Chen X, & Zhou Z. (2023). FUT2 promotes the tumorigenicity and metastasis of colorectal cancer cells via the Wnt/β-catenin pathway. International Journal of Oncology, 62(3), 35.
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2

Splenocyte CD8a Expression Analysis

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Splenocytes were plated into a 48-microwell plate at 1×106 cells per well. The cell suspensions were incubated with fluorochrome-conjugated anti-CD8a monoclonal antibody (eBioscience) and analyzed by FACS Arial (BD Bioscience). The data were analyzed using Flowing Software 2.5.1, Turku, Finland.
Chu X., Li Y., Long Q., Xia Y., Yao Y., Sun W., Huang W., Yang X., Liu C, & Ma Y. (2016). Chimeric HBcAg virus-like particles presenting a HPV 16 E7 epitope significantly suppressed tumor progression through preventive or therapeutic immunization in a TC-1-grafted mouse model. International Journal of Nanomedicine, 11, 2417-2429.
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3

Flow Cytometry Cell Sorting Protocol

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Cells were fixed, labeled with anti H1-HA and anti M2 antibodies and sorted using a FACSArial™ cell sorter (BD biosciences). For details, see SI Appendix, Supplementary Methods.
Chen K.Y., Karuppusamy J., O’Neill M.B., Opuu V., Bahin M., Foulon S., Ibanez P., Quintana-Murci L., Ozawa T., van der Werf S., Nghe P., Naffakh N., Griffiths A, & Isel C. (2023). High-throughput droplet-based analysis of influenza A virus genetic reassortment by single-virus RNA sequencing. Proceedings of the National Academy of Sciences of the United States of America, 120(6), e2211098120.
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4

Single-cell RNA-seq of FACS-sorted Arc+ Neurons

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Tissue was dissociated with FACS lysis buffer (final concentration: 0.32M Sucrose, 10mM Tris − HCl pH8.0, 5mM CaCl2, 3mM Mg(acetate)2, 0.1mM EDTA, 1mM DTT, 0.3% Triton-X −100 and 100× PIC) into single cell suspension, then fixed with 1% formaldehyde for 5 mins, and stop by 0.125M Glycine. Then, cells were washed twice with cold 1xPBS to remove excessed formaldehyde and glycine. After incubating with blocking buffer (Final concentration: 10% normal goat serum, 5% BSA, 0.1% Triton-X-100 and 1XPIC) for half-hour, cells were double-labeled with Arc antibody (Bioss) in 1:20000 dilution per million cell and NeuN antibody (Abcam) in 1:20000 per million cell, together with DAPI (Thermofisher) in 1:2000. PBPT buffer was used for washing (twice each time) and resuspend into 500ul 1x PBS for FACS sorting. FACS was performed on a BD FACSArial (BD Science), and cells were sorted into lysis buffer from Arcturus PicoPure RNA isolation kit (Applied Biosystems). Then, RNA was isolated from sorted cells by Arcturus PicoPure RNA isolation kit (Applied Biosystems) following the manufacturers protocol. 5ng of total RNA per sample and SMRTer Stranded Total RNA-seq Kit v2 Pico Input Mammalian Components kit (Clontech) was used for RNA-seq library preparation following the manufacturers protocol. Sequencing was conducted at GENEWIZ (Suzhou, China) with 150pb Pair-End reads.
Li X., Zhao Q., Wei W., Lin Q., Magnan C., Emami M.R., da Silva L.E., Viola T.W., Marshall P.R., Yin J., Madugalle S.U., Wang Z., Nainar S., Vågbø C.B., Leighton L.J., Zajaczkowski E.L., Ke K., Grassi-Oliveira R., Bjørås M., Baldi P.F., Spitale R.C, & Bredy T.W. (2019). The DNA modification N6-methyl-2’-deoxyadenosine (m6dA) drives activity-induced gene expression and is required for fear extinction. Nature neuroscience, 22(4), 534-544.
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5

Phenotypic Characterization of TERT-BMSCs

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Single cell suspensions (1×106 cells/ml) of TERT-BMSCs, tBMSCs, and BMSCs were prepared in complete DMEM culture medium containing 10% FBS. The cells (300μl) were stained with 5μl of fluorescent conjugated primary antibodies (CD105, 1:200, Clone number: 8A1; CD90, 1:200, Clone number: IBL-6/23; CD44, 1:150, Clone number: T2-F4; CD29, 1:150, Clone number: KM16; CD45, 1:200, Clone number: IBL-3/16; CD34, 1:200, Clone number: ICO-115; CD-11b, 1:200, Clone number: EPR1344, and CD31, 1:200, Clone number: JC/70A, all from Abcam, UK) for 30min at 37°C in the dark. Then, the samples were washed twice with 1ml of PBS buffer and centrifuged at 1000rpm for 5min. The cells were resuspended in 500μl of PBS buffer and detected by a flow cytometer (BD FACSArial.), and analyzed using CellQuest Pro software. We set isotype control as the appropriate control for the median fluorescence intensity by flow cytometry (Supplementary Figure 1 and Supplementary Table 1).
Zhao Y., Chen J., Dai X., Cai H., Ji X., Sheng Y., Liu H., Yang L., Chen Y., Xi D., Sheng M., Xue Y., Shi J., Liu J., Li X, & Dong J. (2017). Human glioma stem-like cells induce malignant transformation of bone marrow mesenchymal stem cells by activating TERT expression. Oncotarget, 8(61), 104418-104429.
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6

Single-cell RNA-seq of FACS-sorted Arc+ Neurons

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Tissue was dissociated with FACS lysis buffer (final concentration: 0.32M Sucrose, 10mM Tris − HCl pH8.0, 5mM CaCl2, 3mM Mg(acetate)2, 0.1mM EDTA, 1mM DTT, 0.3% Triton-X −100 and 100× PIC) into single cell suspension, then fixed with 1% formaldehyde for 5 mins, and stop by 0.125M Glycine. Then, cells were washed twice with cold 1xPBS to remove excessed formaldehyde and glycine. After incubating with blocking buffer (Final concentration: 10% normal goat serum, 5% BSA, 0.1% Triton-X-100 and 1XPIC) for half-hour, cells were double-labeled with Arc antibody (Bioss) in 1:20000 dilution per million cell and NeuN antibody (Abcam) in 1:20000 per million cell, together with DAPI (Thermofisher) in 1:2000. PBPT buffer was used for washing (twice each time) and resuspend into 500ul 1x PBS for FACS sorting. FACS was performed on a BD FACSArial (BD Science), and cells were sorted into lysis buffer from Arcturus PicoPure RNA isolation kit (Applied Biosystems). Then, RNA was isolated from sorted cells by Arcturus PicoPure RNA isolation kit (Applied Biosystems) following the manufacturers protocol. 5ng of total RNA per sample and SMRTer Stranded Total RNA-seq Kit v2 Pico Input Mammalian Components kit (Clontech) was used for RNA-seq library preparation following the manufacturers protocol. Sequencing was conducted at GENEWIZ (Suzhou, China) with 150pb Pair-End reads.
Li X., Zhao Q., Wei W., Lin Q., Magnan C., Emami M.R., da Silva L.E., Viola T.W., Marshall P.R., Yin J., Madugalle S.U., Wang Z., Nainar S., Vågbø C.B., Leighton L.J., Zajaczkowski E.L., Ke K., Grassi-Oliveira R., Bjørås M., Baldi P.F., Spitale R.C, & Bredy T.W. (2019). The DNA modification N6-methyl-2’-deoxyadenosine (m6dA) drives activity-induced gene expression and is required for fear extinction. Nature neuroscience, 22(4), 534-544.
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