CD80 and CD86 expression by monocytes was analyzed by flow cytometry. The samples were processed within 24 h. Erythrocytes lysis was performed using an ammonium chloride solution (0.13 M) and the leukocytes were resuspended with PBS buffer. 106 leukocytes were incubated with PE-conjugated anti-CD80 and FITC-conjugated anti-CD86. The antibodies were diluted 1:100 with PBS and incubated in the dark at 4 °C for 20 min. Cells were analyzed by FACSC Canto II Flow Cytometer (Becton Dickinson, San Jose, CA) with Flow Jo Software (TreeStar). Monocyte cells were identified by manual gating according to side scatter and size.
Facsc canto 2 flow cytometer
Manufactured by BD
Sourced in United States
The FACSC Canto II Flow Cytometer is a laboratory instrument designed for the analysis of cells and particles. It uses the principles of flow cytometry to detect and measure various physical and chemical characteristics of cells as they pass through a laser beam. The device can analyze multiple parameters of individual cells, including size, granularity, and the expression of specific markers on the cell surface or within the cell.
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Lab products found in correlation
4 protocols using facsc canto 2 flow cytometer
1
Monocyte CD80/CD86 Expression Analysis
2
Flow Cytometric Analysis of CD80/CD86 and IL-8 Levels
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CD80 and CD86 expression by lymphocytes and monocytes was analyzed by flow cytometry. The samples were processed within 24 h. Erythrocytes lysis was performed using an ammonium chloride solution (0.13 M) and the leukocytes were resuspended with PBS buffer. 10 6 leukocytes were incubated with PE-conjugated anti-CD80 and FITC-conjugated anti-CD86. The antibodies were diluted 1:100 with PBS and incubated in the dark at 4 °C for 20 min. Cells were analyzed by FACSC Canto II Flow Cytometer (Becton Dickinson, San Jose, CA) with FlowJo Software (TreeStar). Lymphocyte and monocyte cells were identified by manual gating according to side scatter and size.
Serum interleukin-8 (IL-8) levels were quantified by BD TM cytometric bead array (CBA) human inflammation cytokines kit (BD Biosciences, USA) according to the manufacturer's instructions.
Serum interleukin-8 (IL-8) levels were quantified by BD TM cytometric bead array (CBA) human inflammation cytokines kit (BD Biosciences, USA) according to the manufacturer's instructions.
3
Actein Modulates Cell Surface Proteins
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Protein expression on MDA-MB-231 cell surface was studied by flow cytometry. Briefly, 5 × 105 MDA-MB-231 cells in 7 ml culture medium were seeded in 100 mm culture dish and incubated overnight and then treated with actein (10 – 40 μM) for 24 h. After incubation, the attached and floating cells were harvested and washed twice with PBS. Cells were stained with fluorochrome (FITC or Phycoerythrin)-conjugated monoclonal antibodies, CD47b (integrin α2) and CD29 (integrin β1). After incubation at 4°C for 30 min in the dark, cells were washed twice with PBS prior to analysis. The fluorescence of 10,000 events was analyzed by the flow cytometer (FACSC Canto II flow cytometer, BD Biosciences, CA, United States).
4
Cell Cycle Analysis of Breast Cancer Cells
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MDA-MB-361 cells and SK-BR-3 cells (3 × 105/well) were seeded onto a 6-well plate with 3 mL medium and incubated overnight. After 24 h starvation in 1% (v/v) FBS in medium, medium containing 10% (v/v) FBS with actein (10–40 μM) were added into the wells and incubated for 24 or 48 h. Cells were washed with PBS containing 5% (v/v) FBS and ice-cold ethanol (70%) was used to permeabilize cell membrane at 4°C overnight. On the next day, cells were re-suspended in PBS containing RNase A (10 μg/mL) and propidium iodide (20 μg/mL) for 30 min in the dark at 37°C and then analyzed by FACSC Canto II flow cytometer (BD Biosciences, CA, USA).
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