Cells were harvested and incubated with anti-human CD24 phycoerythrin (eBioscience, Gene Company Ltd, Shanghai, China) and anti-human CD44-FITC (eBioscience, Gene Company Ltd) at 4 °C. The cells were then resuspended in 1 ml of phosphate-buffered saline and analysed using a FACScan cytometer (Beckon Dickinson, Oxford, UK).
Anti human cd44 fitc
Manufactured by Thermo Fisher Scientific
Sourced in China, United States
The Anti-human CD44-FITC is a fluorescent-labeled antibody that specifically binds to the human CD44 antigen. CD44 is a cell surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. This product can be used for the identification and enumeration of CD44-positive cells in flow cytometry applications.
Automatically generated - may contain errors
3 protocols using anti human cd44 fitc
1
CD24 and CD44 Cell Phenotyping
2
Cell Cycle and Apoptosis Analysis
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For apoptosis, an Annexin V-FITC apoptosis detection kit (Invitrogen, USA) was used according to the manufacturer's instructions. For cell sorting, after non-specific binding was excluded, approximately 1×106 cells were labeled with conjugated anti-human CD133-PE (eBioscience, USA) and anti-human CD44-FITC (eBioscience, USA). Cells were resuspended in PBS buffer with 2% FBS and analyzed by flow cytometry. Cell sorting was conducted under sterile conditions, and the sorted cells were cultured as described above.
For cell cycle analysis, the harvested cells were suspended in chilled PBS, fixed with cold 70% ethanol and incubated at 4°C for 10 minutes. Then, cells were incubated with 50 μL RNase for 30 min at 37°C. Finally, cells were incubated with propidium iodide for 10 min in the dark at 4°C. The cell cycle distribution was then examined by flow cytometry.
For cell cycle analysis, the harvested cells were suspended in chilled PBS, fixed with cold 70% ethanol and incubated at 4°C for 10 minutes. Then, cells were incubated with 50 μL RNase for 30 min at 37°C. Finally, cells were incubated with propidium iodide for 10 min in the dark at 4°C. The cell cycle distribution was then examined by flow cytometry.
3
Quantifying CD44 and EpCAM Expression
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Transiently transfected KatoIII cells with miR-34a-5p mimic or negative control after 48 h, as well as stably transfected cells were collected by centrifugation at 300×g for 5 min. Antibody staining was performed in 100 µl of phosphate buffer saline (PBS) supplemented with 1% bovine serum albumin (BSA) with the following antibodies: anti-human CD44-FITC (eBioscience, cat.no. 11-0441-81), and anti-human EpCAM-PE (eBioscience, cat.no.12-9326-42). Subsequently, the cells were incubated at 4 °C for 40 min. After being stained, the cells were washed in PBS supplemented with 1% BSA and fixed in 1% formaldehyde. Cells were also treated with appropriate isotype control antibodies (eBioscience). Stained cells were analyzed using flow cytometery. The data were analyzed by FlowJo software 7.6.1 (Tree star, Inc., San Carlos, CA, USA).
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