To analyze expression of cell surface adhesion molecules, 1 × 105 cells were prepared in 0.5% bovine serum albumin (100 μl) and incubated with FcR block (2 μl) for 10 min at room temperature. Thereafter, the cells were washed twice with 1x PBS. Subsequently, the cells were stained with fluorescent-labeled antibodies (1 μl antibody/100 μl cell suspension) and incubated for 30 min at 4 °C. Next, the cells were washed and measured via multicolor flow cytometry. The data were analyzed using FCS express and FlowJo software (Flowjo LLC, Ashland OR) under the institute's license.
Fcs express
Manufactured by FlowJo
Sourced in United States
FCS Express is a data analysis software designed for flow cytometry data. It provides tools for displaying and analyzing flow cytometry data, including creating plots, gating, and statistical analysis.
Automatically generated - may contain errors
4 protocols using fcs express
1
Flow Cytometric Analysis of Cell Surface Adhesion Molecules
2
Cell surface protein expression analysis
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The cell surface expression of ACE2, spike, DR5, FOLR1, etc. was analyzed by flow cytometry in various experiments (see figure and figure legends) after treatment with various control and FuG1 antibodies. After various treatments, cells were trypsinized and suspended in FACS buffer (PBS containing 2% FBS). The single cell suspension was then incubated with primary antibodies for 1 h at 4°C with gentle mixing. Following wash with FACS buffer, the cells were then incubated with fluorescently labeled anti-Rabbit antibody for 1 h. Cells were washed and flow cytometry was performed using FACSCalibur. The data was analyzed by FCS Express (de novo software) and FlowJo.
3
Cell surface protein expression analysis
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The cell surface expression of ACE2, spike, DR5, FOLR1, etc. was analyzed by flow cytometry in various experiments (see figure and figure legends) after treatment with various control and FuG1 antibodies. After various treatments, cells were trypsinized and suspended in FACS buffer (PBS containing 2% FBS). The single cell suspension was then incubated with primary antibodies for 1 h at 4°C with gentle mixing. Following wash with FACS buffer, the cells were then incubated with fluorescently labeled anti-Rabbit antibody for 1 h. Cells were washed and flow cytometry was performed using FACSCalibur. The data was analyzed by FCS Express (de novo software) and FlowJo.
4
Flow Cytometric Analysis of Surface Antigens
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Surface antigens were detected by direct immunofluorescence using flow cytometry (Beckman Coulter, Mississauga, Ontario, Canada and LSRFortessa Cell Analyzer (BD Biosciences, San Jose, California, USA). Briefly, control cells and cells exposed to arsenic were washed twice with PBS supplemented with 5% FBS (Wisent) and 0.01 M sodium azide (flow cytometry buffer). The cells were then exposed to labeled anti-CD29 (eBioscience anti-murine; Pharmingen anti-human: HUTS-21 clone), P-selectin (BD Pharmingen), CD14 (eBioscience), CD41 (BD Pharmingen), Ly6G (eBioscience) or their specific isotype control antibody and incubated for 45 min on ice in the dark. Cells were then washed twice with flow cytometry buffer, fixed in 2% paraformaldehyde and analyzed by flow cytometry. The gates for positive-staining cells were determined by comparison with cells stained with the isotype-matched control antibodies. The FCS express and FlowJo softwares (denovosoftware, CA, USA; flowJo LLC, Ashland, Oregon, USA) were used to analyze the data.
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