1 × 105 cells/ml were fixed and incubated for 15 min at 4 °C with a 1:9 dilution of normal goat serum in PBS. The cells were then labeled with the following antibodies: FITC-conjugated anti-CD44, FITC-conjugated anti-CD34 (Chemicon; Temecula, CA, USA), FITC-conjugated anti-CD45 (Ediscience; USA), PE-conjugated-anti-CD73 (BD; San Jose, CA, USA), PE-conjugated anti-CD90 (Dako, Glostrup, Denmark) and PE-conjugated anti-CD105 (R&D; Minneapolis, MN, USA) for 1 h. The cells were washed and analyzed using a FACS Calibur (BD, NJ) machine and WinMDI software (BD Biosciences).
Pe conjugated anti cd73
Manufactured by BD
Sourced in United States
PE-conjugated anti-CD73 is a monoclonal antibody that binds to the CD73 cell surface protein. CD73 is involved in the regulation of extracellular adenosine levels. The PE conjugate allows for the detection of CD73-expressing cells by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Fitc conjugated anti cd105, by BD (2 mentions)
Fitc conjugated anti cd34, by BD (2 mentions)
Winmdi software, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Fab7335r, by R&D Systems (1 mentions)
Fitc conjugated anti cd31, by R&D Systems (1 mentions)
Facscanto 2, by BD (1 mentions)
Anti cd105, by BD (1 mentions)
Anti cd45, by BD (1 mentions)
Anti cd34, by BD (1 mentions)
Pe conjugated anti cd11b, by BD (1 mentions)
Fitc conjugated anti cd90, by BD (1 mentions)
Fitc conjugated anti cd45, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Percp conjugated anti cd45, by BD (1 mentions)
Fitc conjugated anti cd68, by BioLegend (1 mentions)
Pe conjugated anti cd31, by Thermo Fisher Scientific (1 mentions)
Pe conjugated anti cd34, by BD (1 mentions)
Eclipse ti, by Nikon (1 mentions)
Anti cd44, by Miltenyi Biotec (1 mentions)
6 protocols using pe conjugated anti cd73
1
Stem Cell Phenotype Characterization
2
Isolation and Characterization of OM-MSCs
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Mouse OM-MSCs were obtained from turbinal of wild-type or GFP transgenic mice as previously described [17 ]. After culture expansion in vitro, the isolated OM-MSCs were stained with anti-CD45 (BD, 553079), anti-CD34 (BD, 560238), FITC-conjugated anti-CD31 (R&D, FAB3628G), PE-conjugated anti-CD73 (BD, 550741), anti-CD105 (BD, 562761), and APC-conjugated anti-CD90 (R&D, FAB7335R). Isotype control was used to determine cell gating. One hundred thousand cells/sample were collected using FACS Canto II (BD). We analyzed data using FlowJo V10.5.3 software.
3
Immunophenotyping of Human BM-MSCs
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Cultured human BM-MSC were harvested with TrypLE Select, and 1 × 106 cells were incubated with primary antibodies for 30 min at 4 °C. The primary antibodies used in this study were FITC-conjugated anti-CD105, PE-conjugated anti-CD73, FITC-conjugated anti-CD90, FITC-conjugated anti-CD45, FITC-conjugated anti-CD34, PE-conjugated anti-CD11b, and PE-conjugated anti-CD14, FITC-conjugated anti-CD19 (BD Bioscience). Cells were washed with blocking reagent and analyzed in a FACS Calibur Flow Cytometer (Beckton Dickinson).
4
Immunophenotyping of Mesenchymal Stem Cells
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To fulfill the criteria of the International Society for Cellular Therapy and to exclude contamination of mesenchymal stem cell (MSC) cultures by hematopoietic cells, MSCs were analyzed by flow cytometry. Five antibodies—fluorescein isothiocyanate (FITC)-conjugated anti-CD34, PerCP-conjugated anti-CD45, allophycocyanin(APC)-conjugated anti-CD105, PC7-conjugated anti-CD90, and PE-conjugated anti-CD73 antibodies (BD Biosciences)—were used to identify the MSCs.21 22 (link)
5
Immunophenotyping of Cultured Lung Mesenchymal Cells
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Immunofluorescence staining was performed on cultured LMCs at 24 h after isolation using PE-conjugated anti-CD73 (BD Bioscience, USA, #127205, 1/150), PE-conjugated anti-CD31 (eBioscience, USA, #12-0311-82, 1/150), FITC-conjugated anti-CD68 (BioLegend, #137005, 1/300), and PE-conjugated anti-CD34 (BD Bioscience, #551387, 1/75). Briefly, the cells were washed with PBS and 4% of cold paraformaldehyde was added for 20 min. Then, 5% of goat serum was added for 60 min at RT for blocking, and then, remove without washing. The cells were incubated with the antibodies diluted in BSA/PBS 0.1% for 1–2 h at RT after which the wells were washed with PBS–Tween %0.1, 3 × 5 min. The cells were kept in PBS , and imaging was performed using Nikon Eclipse Ti (Nikon). The ratio of positive cells for each marker was determined by inspecting 100 LMCs.
6
Quantification of Cancer Stem Cell Markers
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HeLa, HCT116 and 22R 1 cells were grown in ultra-low attachment plate for six days with and without treatment of GSK J4 and matrix detached cells were washed two times in ice-cold PBS and resuspended in 100 µl of staining solution (5 µl PE-conjugated anti-CD 73 (BD Bioscience), CD 133 (Miltenyibiotec -130-090-853) and FITC conjugated anti-CD 105(BD Bioscience-56143), and anti-CD44(Miltenyibiotec -130-098-210), 1% FBS, in 1X PBS and incubated at room temperature under the dark condition for 2 hours. Then, cells were washed three times in wash buffer (1%FBS in 1X PBS) and then analyzed by Guava Easy-Cyte ow cytometer. For all assays, 10,000 cells were taken for measurement and also for analysis. For each plot 1000 cells were displayed.
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