Cytexpert 1

Manufactured by Beckman Coulter

Sourced in United States

The CytExpert 1.1 is a flow cytometry instrument designed for cell analysis. The core function of the CytExpert 1.1 is to detect and measure the physical and chemical characteristics of cells or other particles in a fluid as they flow in a single file through a beam of light.

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Lab products found in correlation

Cytoflex, by Beckman Coulter (11 mentions) Ionomycin, by Merck Group (6 mentions) Phorbol myristate acetate (pma), by Merck Group (5 mentions) Fixation and permeabilization solution, by BD (4 mentions) Brefeldin a bfa, by Merck Group (4 mentions) Saponin, by Merck Group (4 mentions) Beckman cytoflex, by Beckman Coulter (3 mentions) Brefeldin a, by Merck Group (2 mentions) Pe conjugated anti human bcl2, by BD (2 mentions) Flow cytometric analysis, by Bio-Rad (2 mentions) Isotype control, by Thermo Fisher Scientific (1 mentions) Flow cytometry, by Beckman Coulter (1 mentions) Cytoflex instrument, by Beckman Coulter (1 mentions) Apoptosis detection kit, by BD (1 mentions) Rabbit anti human sirt1, by Santa Cruz Biotechnology (1 mentions) Cytoflex flow cytometer, by Beckman Coulter (1 mentions) Annexin 5 pe, by BD (1 mentions) 7 aminoactinomycin d, by BD (1 mentions) Annexin 5 propidium iodide, by BD (1 mentions) Gr 1 rb6 8c5, by BD (1 mentions)

Spelling variants of this product

CytExpert (205 citations) CytExpert for DxFLEX version 1.0 (2 citations)

26 protocols using cytexpert 1

Multiparametric Flow Cytometry for Immune Cell Profiling

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Cells were washed twice with PBS and blocked in PBS buffer containing 1% BSA for 30 min. Cells were then stained with speci c antibodies for the cell surface antigens for 30 min at 4°C in the dark. The phenotypes (1 × 10 6 cells per run) were analyzed using ow cytometry (Beckman CytoFLEX) and CytExpert 1.1 (Beckman Coulter Inc.). The single nuclear cells were gated to exclude the dead cells and doublet. Isotype-matched controls for each cell surface marker were included in the staining protocol.
Cell intracellular cytokine staining 1.5 × 10 6 cells were resuspended in complete RPMI 1640 medium, then stimulated with phorbol 12myristate 13-acetate (PMA) (20 ng/ml, Sigma) and ionomycin (1 μg/ml, Sigma) for 1 h. Brefeldin A (BFA, 10 μg/ml, Sigma) was added and incubated for 4 h. Cells were washed twice in PBS and stained with speci c antibodies for the cell surface antigens for 30 min at 4°C in the dark. Cells were xed with Fixation and Permeabilization Solution (BD Biosciences) for 20 min at 4°C in the dark. Next, cells were stained with speci c antibodies for each cytokine. The results were analyzed using ow cytometry (Beckman CytoFLEX) and CytExpert 1.1 (Beckman Coulter Inc.). The single nuclear cells were gated to exclude the dead cells and doublet. Isotype-matched controls for each cell surface marker were performed in the staining protocol.

Wei H., Jin C., Peng A., Xie H., Xie S., Feng Y., Xie A., Li J., Fang C., Yang Q., Qiu H., Qi Y., Yin Z., Wang X., & Huang J. (2020). Characterization of γδT Cells in Lung of Plasmodium yoelii-infected C57BL/6 Mice.

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Comprehensive Cytokine Profiling of Activated T Cells

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1.5 × 10⁶ cells were resuspended in complete RPMI 1640 medium, then stimulated with phorbol 12-myristate 13-acetate (PMA) (20 ng/ml, Sigma) and ionomycin (1 µg/ml, Sigma) for 1 h. Brefeldin A (BFA, 10 µg/ml, Sigma) was added and incubated for 4 h. Cells were washed twice in PBS and stained with specific antibodies for the cell surface antigens for 30 min at 4 °C in the dark. Cells were fixed with Fixation and Permeabilization Solution (BD Biosciences) for 20 min at 4 °C in the dark. Next, cells were stained with specific antibodies for each cytokine. The results were analysed using flow cytometry (Beckman CytoFLEX) and CytExpert 1.1 (Beckman Coulter Inc.). The single nuclear cells were gated to exclude the dead cells and doublet. For gating CD3⁺ γδTCR⁺ cells, CD3⁺, CD3⁺ CD4⁺, CD3⁺ CD8⁺, CD3⁻ CD19⁺ cells, FMO controls were used. Isotype controls were used for intracellular cytokines staining. 1,500,000 cells were used for cell intracellular cytokine staining, and 300,000 events were collected for each tube.

Wei H., Jin C., Peng A., Xie H., Xie S., Feng Y., Xie A., Li J., Fang C., Yang Q., Qiu H., Qi Y., Yin Z., Wang X, & Huang J. (2021). Characterization of γδT cells in lung of Plasmodium yoelii-infected C57BL/6 mice. Malaria Journal, 20, 89.

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Immunophenotyping of Lymphocyte Subsets

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Cells were washed twice with PBS and blocked in PBS buffer containing 1% BSA for 30 min. Cells were then stained with specific antibodies for the cell surface antigens for 30 min at 4 °C in the dark. The phenotypes (1 × 10⁶ cells per run) were analysed using flow cytometry (Beckman CytoFLEX) and CytExpert 1.1 (Beckman Coulter Inc.). The single nuclear cells were gated to exclude the dead cells and doublet. For gating CD3⁺ γδTCR⁺ cells, CD3⁺, CD3⁺ CD4⁺, CD3⁺ CD8⁺, CD3⁻ CD19⁺ cells, fluorescence minus one (FMO) controls were used. For other surface makers, isotype controls were used. 1,000,000 cells were used for cell surface staining, and 300,000 events were collected for each tube.

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Flow Cytometric Analysis of Cell Surface and Intracellular TLR3 Expression

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Cells were then stained with conjugated antibodies specific for the cell surface antigens for 30 min. Then, expressions phenotypes of antibody-labeled cells were analyzed by flow cytometry (Beckman CytoFLEX), and the results were analyzed using the CytExpert 1.1 (Beckman Coulter Inc.). Isotype-matched controls for cell surface markers were included in each staining protocol. When the samples were stained for TLR3, cells were fixed with fixation and permeabilization solution (BD Biosciences) for 20 min at 4 °C in the dark. Then, cells were permeabilized by PBS buffer containing 0.1% saponin (Sigma), 1% BSA, and 0.05% NaN₃ overnight at 4 °C. Next, cells were stained with conjugated antibodies that were specific for TLR3.

Xie H., Chen D., Feng Y., Mo F., Liu L., Xing J., Xiao W., Gong Y., Tang S., Tan Z., Liang G., Zhao S., Yin W, & Huang J. (2024). Evaluation of the TLR3 involvement during Schistosoma japonicum-induced pathology. BMC Immunology, 25, 2.

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Cytokine Profiling of Lymphocytes

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Single lymphocyte suspensions were isolated from the lung, and the cell concentration was adjusted to 1.5 × 10⁶/ml. Cells were then stimulated with phorbol 12-myristate 13-acetate (PMA) (20 ng/ml, Sigma) and ionomycin (1 μg/ml, Sigma) for 5 h (37°C, 5% CO₂). Brefeldin A (BFA, 10 μg/ml, Sigma) was added during the last 4 h of incubation. Cells were washed twice in PBS and stained for 30 min at 4°C in the dark with conjugated antibodies specific for the cell surface antigens CD3 and γδ TCR. Cells were fixed with Fixation and Permeabilization Solution (BD Biosciences) for 20 min at 4°C in the dark and permeabilized overnight at 4°C in PBS buffer containing 0.1% saponin (Sigma), 1% BSA, and 0.05% NaN₃. Next, cells were stained with conjugated antibodies that specific for each cytokine. Expression phenotypes of antibody-labeled lymphocytes (1.5 × 10⁶ cells per run) were analyzed using flow cytometry (Beckman CytoFLEX), and the results were analyzed using CytExpert 1.1 (Beckman Coulter Inc.). The region of single nuclear cells was gated to ensure the dead cell and doublet exclusion. Isotype-matched controls for cytokines were included in each staining protocol.

Cha H., Xie H., Jin C., Feng Y., Xie S., Xie A., Yang Q., Qi Y., Qiu H., Wu Q., Yin Z., Mu J, & Huang J. (2020). Adjustments of γδ T Cells in the Lung of Schistosoma japonicum-Infected C56BL/6 Mice. Frontiers in Immunology, 11, 1045.

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Splenic Lymphocyte Profiling by Flow Cytometry

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Splenic lymphocyte stimulation was performed as previously described [7 (link)]. Briefly, for cell surface staining, single splenic lymphocyte suspensions were washed twice and stained with anti-CD3, NK1.1, TLR2, TLR4, CD8, CD4, γδT, CD19, Ly6G, F4/80, MHC II, CD69, CD94 (NKG2A/C/E), or CD314 (NKG2D) antibodies for 30 min at 4°C in the dark. Stained cells were washed twice and detected by using flow cytometry. For intracellular cytokine staining, single splenic lymphocyte suspensions were incubated for 5 h in the presence of phorbol 12-myristate 13-acetate (PMA) (20 ng/ml, Sigma) and ionomycin (1 μg/ml, Sigma) at 37°C under a 5% CO₂ atmosphere; brefeldin A (10 μg/ml, Sigma) was added 1 hour after stimulation. Cells were washed twice and stained with antibodies of cell surface markers for 30 min at 4°C in the dark. Cells were fixed with 4% paraformaldehyde and permeabilized overnight at 4°C in PBS buffers containing 0.1% saponin (Sigma), 1% BSA, and 0.05% NaN₃. In the next day, cells were stained with conjugated antibodies specific for TLR3, TLR7, IL-4, IFN-γ, IL-17, and IL-5 for 30 min. Stained cells were washed twice and detected by using flow cytometry (CytoFLEX, Beckman Coulter, USA), and data were analyzed by the program CytExpert 1.1 (Beckman Coulter, USA).

Qu J., Li L., Xie H., Zhang X., Yang Q., Qiu H., Feng Y., Jin C., Dong N, & Huang J. (2018). TLR3 Modulates the Response of NK Cells against Schistosoma japonicum. Journal of Immunology Research, 2018, 7519856.

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CD4+ T Cell Apoptosis Analysis

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CD4⁺T lymphocyte apoptosis was detected using the Annexin V, 7-aminoactinomycin D (7AAD) and anti-mouse CD4 staining method. Briefly, after 3 days of BMMSCs and CD4⁺T lymphocyte co-culture, supernatants were collected and centrifuged for 5 min at 400 × g (4°C). The resultant cell pellets were collected, washed and dispersed in PBS solution containing 3% FBS to a concentration of 1×10⁶ cells/ml. Subsequently, fluorescent anti-mouse CD4 antibody (1:50; BioLegend, Inc., cat. no. 100413) was added and incubated at 4°C in the dark for 1 h. AnnexinV (5 µl) and 7AAD (5 µl) were subsequently added and the whole system was incubated in the dark at room temperature for 15 min. Finally, the apoptotic ratio of CD4⁺T lymphocytes was determined by flow cytometry and analyzed by equipped software (CytExpert1.1, Beckman Coulter, Inc.).

Shao B., Fu X., Yu Y, & Yang D. (2018). Regulatory effects of miRNA-181a on FasL expression in bone marrow mesenchymal stem cells and its effect on CD4+T lymphocyte apoptosis in estrogen deficiency-induced osteoporosis. Molecular Medicine Reports, 18(1), 920-930.

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Lymphocyte Immunophenotyping by Flow Cytometry

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Cells were washed in PBS and blocked in PBS buffer containing 1% BSA for 30 min. Cells were then stained for 30 min at 4°C in the dark with conjugated antibodies specific for the cell surface antigens. Expression phenotypes of antibody-labeled lymphocytes (1 × 10⁶ cells per run) were analyzed using flow cytometry (Beckman CytoFLEX), and the results were analyzed using the CytExpert 1.1 (Beckman Coulter Inc.). The region of single nuclear cells was gated to ensure the dead cell and doublet exclusion. Isotype-matched controls for cell surface markers were included in each staining protocol.

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Multiparameter Immune Cell Profiling

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Cells were washed twice with PBS and blocked in PBS buffer containing 1% BSA for 30 min. Cells were then stained for 30 min at 4 °C in the dark with conjugated antibodies specific for the cell surface antigens CD3, CD4, CD8, γδ T, CD25, CD44, CD69,Vγ2, CD62L, CD40L, CD16/32, and PD-1. Cells were analyzed using a flow cytometer (Beckman CytoFLEX), and the results were analyzed using CytExpert 1.1 software (Beckman Coulter, Inc.). Isotype-matched controls for cytokines were included in each staining protocol.

Xie H., Xie S., Wang M., Wei H., Huang H., Xie A., Li J., Fang C., Shi F., Yang Q., Qi Y., Yin Z., Wang X, & Huang J. (2022). Properties and Roles of γδT Cells in Plasmodium yoelii nigeriensis NSM Infected C57BL/6 Mice. Frontiers in Cellular and Infection Microbiology, 11, 788546.

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Multiparameter Lymphocyte Profiling

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For cell surface marker detection, single lymphocytes were stained with fluorescence-labeled anti-PD-1, CD11b, CD3, CD19, CD4, CD8, CD62L, CD69, CD40L, CD25, and CXCR5 antibodies at 4°C for 30 min. For cytokine detection, single lymphocytes were stimulated with phorbol 12-myristate 13-acetate (PMA, 20 ng/ml, Sigma, Germany) and ionomycin (1 μg/ml, Sigma) at 37°C for 1 h, then brefeldin A (10 μg/ml, Sigma, Germany) was used to stop the stimulation. Cells were incubated with antibodies for cell surface markers firstly, then they were fixed with 4% paraformaldehyde, and permeabilized. The cells were incubated with fluorescence-labeled antibodies for IFN-γ, IL-10, IL-17, IL-4, and IL-2 for 30 min. To detect the expression of NFATc1 and HIF-1α, cells were treated with a transcription factor staining kit (00-5523-00, Invitrogen,United States), and incubated with fluorescent antibodies for NFATc1 and HIF-1α. Stained cells were determined by FCM and the results were analyzed by CytExpert 1.1 (Beckman coulter, United States).

Wei H., Xie A., Li J., Fang C., Liu L., Xing J., Shi F., Mo F., Chen D., Xie H., Yang Q., Pan X., Tang X, & Huang J. (2022). PD-1+ CD4 T cell immune response is mediated by HIF-1α/NFATc1 pathway after P. yoelii infection. Frontiers in Immunology, 13, 942862.

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