BMSCs were incubated with FITC conjugated CD44, PE-conjugated CD105, PerCP-Cy 5.5-conjugated CD34, and PE-Cy7-conjugated CD45 (10 μg/ml: BD Biosciences) for 20 min at room temperature, which were further fixed with 2% formaldehyde/ phosphate-buffered saline (PBS) solution on ice. Isotype controls were also utilized to confirm the specific staining. At least 50,000 singlet events were detected on a BD Facscanto II Flow Cytometer.
Fitc conjugated cd44
Manufactured by BD
Sourced in United States
FITC-conjugated CD44 is a fluorescently labeled cell surface marker for the detection and identification of CD44-positive cells. CD44 is a cell adhesion molecule involved in various cellular processes. This product provides a tool for the analysis of CD44-expressing cells using flow cytometry or other fluorescence-based applications.
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Lab products found in correlation
Pe conjugated cd105, by BD (1 mentions)
Pe cy7 conjugated cd45, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Pe conjugated cd24, by BD (1 mentions)
Accuri c6, by BD (1 mentions)
Magnetic cell sorting system, by Miltenyi Biotec (1 mentions)
Bd555742, by BD (1 mentions)
Fitc conjugated cd44 antibody, by BD (1 mentions)
Accutase, by Innovative Cell Technologies (1 mentions)
Facscalibur, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Fitc mouse igg2b κ isotype control, by BD (1 mentions)
Annexin 5 fitc propidium iodide apoptosis detection kit, by BD (1 mentions)
Cycletest plus dna reagent kit, by BD (1 mentions)
5 protocols using fitc conjugated cd44
1
Multicolor Flow Cytometry Analysis of BMSCs
2
Breast Cancer Cell Immunophenotyping
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We followed a previously described method [39 (link)]. A total of 1 × 106 cells were incubated with FITC-conjugated CD44 and PE-conjugated CD24 (BD) and incubated at 4 °C for 20 min. The breast cancer cells were washed twice with 1XPBS and then analyzed by using a flow cytometer (Accuri C6, BD).
3
Cell Sorting and Quantification via CD44
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For cell sorting, cells were dissociated using Accutase (Innovative Cell Technologies). For FACS, cells were resuspended in PBS containing 0.5% BSA, stained with FITC-conjugated CD44 (BD555478) or isotype control antibody (BD555742) from BD Biosciences and analysed on a BD FACSCalibur (BD Biosciences) using Cell Quest software. CD44-positive cells were sorted using a magnetic cell sorting system (Miltenyi Biotech). Cells were stained with CD44 Micro-Beads, passed through a LS magnetic column, then eluted from the column after removal from the magnet. CD44-positive cells were quantified by immunofluorescence using FITC-conjugated CD44 antibody (555478; BD Biosciences).
4
Mesenchymal Stem Cell Surface Markers
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To identify the characteristics of mesenchymal stem cells, surface marker protein expression was evaluated by flow cytometry. hDPSCs and SCAPs were stained with FITC-conjugated CD44 (BD Bioscience Pharmingen, United States), FITC-conjugated CD73 (BD Bioscience Pharmingen, United States), PE-conjugated CD105 (Immuno Tools, Germany), APC-conjugated CD90 (Immuno Tools, Germany), and PerCP-conjugated CD45 antibodies (Immuno Tools, Germany) (1:50 dilution for all antibodies). Mouse IgG isotype was used as the control. The stained cells were analyzed by FACSCalibur Flow Cytometer (Becton Dickinson, Worldwide Inc, United States).
5
Multiparametric Flow Cytometry Analysis
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For the cell cycle assay, cells were collected and fixed in ice-cold 75% ethanol overnight. The cells were then incubated with 500 μL of propidium iodide (PI) staining solution for 30 min using a Cycletest Plus DNA Reagent Kit (BD Biosciences). The cell distribution was detected by flow cytometry.
For apoptosis analysis, cells were stained with an Annexin V-FITC/Propidium Iodide (PI) Apoptosis Detection Kit (BD, Biosciences #556547) and detected by flow cytometry.
For detection of the CD44 proportion, cells were resuspended in PBS containing 2% FBS and incubated with FITC-conjugated CD44 (BD Biosciences, 555478) for 20 min. After incubation, the cells were washed and resuspended in 300 μl of PBS and analyzed using flow cytometry. FITC Mouse IgG2b κ Isotype Control (BD Biosciences, 556655) was used as a control.
For apoptosis analysis, cells were stained with an Annexin V-FITC/Propidium Iodide (PI) Apoptosis Detection Kit (BD, Biosciences #556547) and detected by flow cytometry.
For detection of the CD44 proportion, cells were resuspended in PBS containing 2% FBS and incubated with FITC-conjugated CD44 (BD Biosciences, 555478) for 20 min. After incubation, the cells were washed and resuspended in 300 μl of PBS and analyzed using flow cytometry. FITC Mouse IgG2b κ Isotype Control (BD Biosciences, 556655) was used as a control.
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