Antibodies against anti-human CD73-PE, CD90-PE, CD105-PE, CD34-FITC, and CD45-PE (Becton Dickinson, San Jose, CA, USA) were used at 1:100 dilution. Cells were analyzed using FACSCalibur™ (Becton Dickinson).
Cd105 pe
Manufactured by BD
Sourced in United States
CD105-PE is a laboratory reagent used for the detection and quantification of CD105 (Endoglin) expression on cells. It is a monoclonal antibody conjugated with the fluorescent dye Phycoerythrin (PE), which allows for the identification and analysis of CD105-positive cells using flow cytometry or other fluorescence-based techniques. The core function of CD105-PE is to provide a reliable and specific tool for researchers to study the expression of this cell surface marker.
Automatically generated - may contain errors
Lab products found in correlation
Cd73 pe, by BD (35 mentions)
Cd45 fitc, by BD (25 mentions)
Cd90 fitc, by BD (25 mentions)
Cd90 pe, by BD (24 mentions)
Cd34 pe, by BD (22 mentions)
Cd34 fitc, by BD (19 mentions)
Cd45 pe, by BD (16 mentions)
Cd44 pe, by BD (15 mentions)
Cd29 pe, by BD (12 mentions)
Facscalibur, by BD (11 mentions)
Cd14 fitc, by BD (11 mentions)
Cd73 fitc, by BD (10 mentions)
Hla dr fitc, by BD (10 mentions)
Cd90 apc, by BD (10 mentions)
Cd31 pe, by BD (9 mentions)
Hla dr pe, by BD (9 mentions)
Cellquest software, by BD (8 mentions)
Cd14 pe, by BD (8 mentions)
Facscalibur flow cytometer, by BD (8 mentions)
Cd45 apc, by BD (8 mentions)
93 protocols using cd105 pe
1
Flow Cytometry Immunophenotyping of Cells
2
Optimized hASCs Culture Protocol
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hASCs and its culture media components were purchased from Guangzhou Cyagen Biosciences co., LTD (Guangzhou, China). Penicillin–streptomycin, fetal bovine serum (FBS), Dulbecco's Phosphate Buffered Saline (D-PBS), and Trypsin were purchased from Gibco (USA). CD105-PE, CD73-PE, HLA-ABC-PE, CD14-FITC, CD34-PE, and CD45-PE antibody were purchased from Becton–Dickinson (USA). Trizol reagent was purchased from Life Technologies (USA). MiniBEST Universal RNA Extraction Kit, SYBR Premix Ex Taq II (Tli RNaseH Plus) kit, and PrimeScript RT Master Mix kit (Perfect Real Time) were purchased from TaKaRa (Japan). RNeasy Mini Kit was purchased from Qiagen (Germany). Cell cycle kit and Annexin V-FITC/PI apoptosis kit were purchased from Multisciences (Lianke) Biotech, co., LTD (Hangzhou, China). Cell Counting Kit-8 (CCK-8) kits were purchased from Dojindo (Osaka, Japan). Ki-67 cell proliferation kit (IF) was purchased from Sangon Biotech (Shanghai) Co., Ltd (Shanghai, China). Enzyme-Linked Immunosorbent assay kit was purchased from Abcam (United Kingdom). The Scepter 2.0 cell counter was purchased from Merck Millipore (Massachusetts, USA). The chromatographic column was purchased from Waters (USA). Acetonitrile and methanol were purchased from Merck (Darmstadt, Germany). Formic acid was purchased from CNW Technologies (Shanghai, China).
3
Characterization of BM-MSCs Prior to Infusion
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BM‐MSCs were characterized just before infusion, by cell surface expression of CD73‐PE, CD90‐PE, CD19‐PE, CD34‐PE, and CD45‐PE (BioLegend, San Diego, CA); CD105‐PE, HLADR‐PE, and CD14‐PE (Becton Dickinson); propidium iodide (PI; Sigma); and isotype IgG1‐PE and isotype IgG2a‐PE (Becton Dickinson). For immunolabeling, cells were incubated for 45 minutes at 2°C–8°C. PI‐negative BM‐MSCs (viable) were characterized using FC500 flow cytometer (Beckman Coulter, Mississauga, ON, Canada) and analyzed using FlowJo (Treestar).
4
Osteogenicity of Orbicularis Oris MSCs
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The osteogenicity of MSCs from the orbicularis oris muscle has been studied by our laboratory and has been previously shown to induce in vitro and in vivo bone formation in critical-sized calvarial osseous defects in rats.20 (link)Flow cytometry analysis was performed using a Guava Easy Cyte microcapillary flow cytometer (Guava Technologies, Hayward, CA, USA) utilizing laser excitation and emission wavelengths of 488 and 532 nm. Cells were pelleted and resuspended in phosphate-buffered saline (PBS; Gibco-Invitrogen®, Carlsbad, CA, USA) at a concentration of 1.0 × 106 cells/mL and stained with saturating concentrations of antibodies. After a 45-min incubation in the dark at room temperature, cells were washed three times with PBS and resuspended in 0.25 mL of cold PBS. In order to analyze cell surface expression of typical protein markers, adherent cells were treated with the following primary anti-human antibodies: CD29-PE-Cy5, CD31-PE, CD45-FITC, CD73-PE, CD90-R-PE, and CD105-PE (Becton Dickinson, Franklin Lakes, NJ, USA). Unstained cells were gated on forward scatter to eliminate particulate debris and clumped cells. A minimum of 5000 events were counted for each sample.
These cells presented high expression of MSC markers (CD73-PE, CD90-R-PE, CD105-PE) and lack of expression of hematopoietic and endothelial markers (CD29-PE-Cy5, CD31-PE, CD45-FITC).
These cells presented high expression of MSC markers (CD73-PE, CD90-R-PE, CD105-PE) and lack of expression of hematopoietic and endothelial markers (CD29-PE-Cy5, CD31-PE, CD45-FITC).
5
Immunophenotyping of IVD Progenitor Cells
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IVD progenitors were analyzed for the expression of mesenchymal and hematopoietic surface marker molecules, by direct immunofluorescent staining, as previously reported [10 (link)]. Briefly, cell pellets were resuspended in phosphate buffered solution (PBS), and incubated with fluorescein isothiocyanate (FITC)– or phycoerythrin (PE)–conjugated mouse anti-human antibodies CD14-FITC (#F0844; DakoCytomation; Dako, Glostrup, Denmark), CD44-PE, and CD105-PE (#550989, #560839; Becton Dickinson, Franklin Lakes, NJ, USA) for 15 min at 4 °C. Monoclonal antibodies with no specificity were used as negative control. Antibody-treated cells were then washed with PBS and spun down (300 xg). Cell pellets were resuspended in 400 μL of PBS and analyzed by FACS Scan (Becton Dickinson). For each sample, 20000 events were acquired and analyzed using the CellQuest software (Becton Dickinson).
6
Characterization of DPSC Cell Populations
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DPSC strains were characterized using flow cytometry. One hundred thousand cells (1 × 105) from each population were used. The cells were stained with the following monoclonal antibodies: CD29-PE, CD31-FITC, CD34-FITC, CD44-PE, CD45-PE, CD73-FITC, CD90-FITC, CD105-PE and CD117-PE (Becton Dickinson, Franklin Lakes, NJ, USA). Appropriate isotype-matched control antibodies were used for all of the analyses. Cells were analyzed using a FACSCalibur flow cytometer with Cell Quest Software (Becton Dickinson).
7
Mesenchymal Origin Verification of AD-MSCs
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In order to confirm the mesenchymal origin of the obtained AD-MSC lineages, cell membrane protein expression analysis was performed by flow cytometry, using the methods previously described by the present group.11 (link), 12 (link), 13 (link), 14 (link) To that end, the adherent cells obtained in stage five were incubated with the following conjugated antibodies: CD29-PE-Cy5, CD31-PE, CD34-PerCP, CD45-FITC, CD73-PE, CD90-R-PE, CD105-PE, HLA-ABC-FITC, HLA-DR-R-PE (Becton Dickinson), according to the manufacturer's recommendations. As a negative control, cells were incubated with PBS instead of the primary antibody. A total of 10,000 events were acquired with the Guava EasyCyte flow cytometer (Guava Technologies) and analyzed with Guava ExpressPro software (Guava Technologies).
8
Phenotypic Characterization of Mesenchymal Stem Cells
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HBM-MSCs at passage 3 were trypsinized, centrifuged at 300 ×g for 8 min, and resuspended in PBS at a concentration of 1 × 106 cells/mL. Aliquots of 100 μL were incubated for 30 min in 20 μL of antibodies against CD14, CD45 (FITC) or CD73, CD34 phycoerythrin (PE) or in 5 μL of CD105 PE or CD90 (FITC) (Becton, Dickinson, United States), washed with 1 mL of stain buffer (BD-Pharmingen, United States), and resuspended in 500 μL of stain buffer. The labeled cells were analyzed using an argon ion laser at a wavelength of 488 nm (FACSCalibur, Becton, Dickinson, United States). A total of ten thousand events were obtained and analyzed using CellQuest software (Becton, Dickinson, United States). Control staining using the appropriate isotype-matched monoclonal antibodies was included.
9
Mesenchymal Stem Cell Immunophenotyping
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To evaluate the MSC immunophenotype a flow cytometer analysis was performed at passage 4. After the detachment, cells where washed with Dulbecco’s phosphate buffered saline (DPBS 1× without Ca2+/Mg2+; Euroclone) and briefly incubated with the following conjugated monoclonal antibodies and isotype controls for 15 min at room temperature: CD90 FITC, CD105 PE, CD73 PE, HLA-DR PE-Cy7, CD45 APC-Cy7, CD14 APC-Cy7, CD19 PE-Cy7,CD34 APC (Becton Dickinson), APC-Cy7 IGg1 isotype control, APC-Cy7 IGg2 isotype control, PE-Cy7 IGg1 isotype control, PE-Cy7 IGg2 isotype control, FITC IGg1 isotype control, PE IGg1 isotype control and APC IGg1 isotype control (BD Pharmingen). The vitality dye 7-aminoactinomycin D (7-AAD; BD Pharmingen) was also added to discriminate dead cells during flow cytometry analysis. After incubation, cells were washed, resuspended in DPBS and then analyzed by flow cytometry on a FACS Canto II (Becton Dickinson) equipped with the DIVA software program.
10
Surface Marker Expression of Adipose-Derived Stem Cells
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The expression of ASCs characteristic surface markers was analyzed by flow cytometry technique, for abdominal and breast ASCs, as previously described (Fathi and Farahzadi 2018 (link); Fathi et al. 2019 (link)) with some modifications. At the second or third passage, a total of 5 × 104 cells were harvested, enzymatically using 0.25% Trypsin-EDTA (Lonza) or manually using a cell scraper, centrifuged at 1600 rpm, and then washed with PBS (Lonza). Cells were incubated for 20–30 min in the dark with the suitable volumes of the following fluorochrome-labeled monoclonal antibodies: CD90-FITC, CD105-PE, CD44-PE, and CD34-FITC (BD, Biosciences, USA). After incubation, another washing step was done using PBS to remove excess stain. Flow cytometric analysis was performed using a Coulter EPICS device (Beckman Coulter, USA) and data analysis was carried out using System II Software.
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