Surface markers for hADSCs were evaluated using a mini Guava EasyCyte flow cytometer. 5x104 cells were incubated with fluorescein isothiocyanate (FITC) or phycoerythrin (PE)-conjugated antibodies, anti-CD105, anti-CD34, anti-STROI, anti-CD73, anti-CD45, anti-HLA-ABC, anti-HLA-DR (Invitrogen, Frederick, MD) for 30 min at 4°C in PBS and washed afterwards. Cell autofluorescence in channel F1 or F2 was subtracted to obtain a neat signal of each marker. 5x104 cells were plated for differentiation tests. Adipogenic differentiation was performed in StemPro adipogenesis-conditioned medium (Invitrogen, Grand Island, NY) for one week, replacing medium every 3 days. Lipid droplets were stained by using red oil staining for validating differentiation. StemPro osteogenesis-conditioned medium (Invitrogen, Grand Island, NY) was used for osteogenic differentiation for 11 days, replacing media every 48 hours. Extracellular calcium deposits were identified by Von Kossa staining. hADSCs cultured in DMEN were also stained as controls.
Anti cd105
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
Anti-CD105 is a laboratory reagent used for the detection and identification of CD105-positive cells. CD105, also known as endoglin, is a transmembrane glycoprotein expressed on endothelial cells and is involved in angiogenesis. This reagent can be used in various research applications, such as flow cytometry and immunohistochemistry, to study the expression of CD105 in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd45, by Thermo Fisher Scientific (7 mentions)
Anti cd73, by Thermo Fisher Scientific (6 mentions)
Anti cd90, by Thermo Fisher Scientific (6 mentions)
Anti cd44, by Thermo Fisher Scientific (6 mentions)
Anti cd90, by BD (6 mentions)
Anti cd34, by Thermo Fisher Scientific (5 mentions)
Anti cd31, by Thermo Fisher Scientific (5 mentions)
Anti cd29, by Thermo Fisher Scientific (4 mentions)
Facsdiva software, by BD (3 mentions)
Anti cd45, by BD (2 mentions)
Anti cd34, by BD (2 mentions)
Anti cd31, by BD (2 mentions)
Anti cd146, by Thermo Fisher Scientific (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Anti cd90, by BioLegend (2 mentions)
Facscanto 2, by BD (2 mentions)
Anti hla dr, by Thermo Fisher Scientific (1 mentions)
Anti hla abc, by Thermo Fisher Scientific (1 mentions)
Nestin, by Thermo Fisher Scientific (1 mentions)
Anti f4 80, by Thermo Fisher Scientific (1 mentions)
24 protocols using anti cd105
1
Characterization and Differentiation of hADSCs
2
Isolation and Characterization of PDLSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fourth-generation PDLSCs were collected and washed twice with PBS containing 1% FBS. After incubation in the dark with anti-CD45, anti-CD31, anti-CD29, anti-CD90 and anti-CD105 antibodies (Invitrogen, California, USA) at 4 °C for 30 minutes. The cells were centrifuged and washed three times, and the suspension was analyzed by sorted flow cytometry (Influx, BD, New Jersey, USA). The detection of DPSC surface antibodies was the same as above, and the markers included anti-CD73, anti-CD90, anti-CD45, anti-CD31 and Nestin (Invitrogen, California, USA).
Macrophage polarization markers were detected as follows. After washing with PBS, centrifuged cells with anti-F4/80 and anti-CD86 or anti-CD206 (Invitrogen, California, USA) were incubated in the dark at 4 °C for 30 minutes. The cells were then washed once, suspended in PBS containing 1% FBS and analyzed by flow cytometry. Flow cytometry data were analyzed by FlowJo software v10.2 (BD, New Jersey, USA).
Macrophage polarization markers were detected as follows. After washing with PBS, centrifuged cells with anti-F4/80 and anti-CD86 or anti-CD206 (Invitrogen, California, USA) were incubated in the dark at 4 °C for 30 minutes. The cells were then washed once, suspended in PBS containing 1% FBS and analyzed by flow cytometry. Flow cytometry data were analyzed by FlowJo software v10.2 (BD, New Jersey, USA).
3
Immunohistochemical Analysis of Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical analysis of paraffin-embedded specimens was performed as previously described [29 (link), 37 (link), 38 (link)]. Antibodies anti-ΔNp63 (anti-p40; Calbiochem, Gibbstown, NJ, USA), anti-HβD1 (Biologo, Kronshagen, Germany), anti-HβD2 (Abcam, Cambridge, MA, USA), anti-HβD4 (Abcam), anti-CD105 (Thermo Scientific, Waltham, MA, USA), anti-Podoplanin (Clone D2-40, Dako, Glostrup, Denmark), anti-alpha Smooth muscle actin (SMA) (Abcam) and anti-Prox1 (ReliaTech GmbH, Wolfenbuettel, Germany) were used for the primary reaction. Immunoperoxidase staining was performed using the Envision kit (Dako, Glostrup, Denmark) or the BrightVision Plus kit (Immunologic, Duiven, Netherlands) according to the supplier's recommendations. Positive cells were visualized using a 3, 3'-diaminobenzidine (DAB) substrate and the sections were counterstained with hematoxylin. A control IgG was used as negative control (Santa Cruz Biotechnology, Santa Cruz, CA, USA). To test the specificity of the HβD staining, the different anti-HβD antibodies were neutralized by the incubation with an excess of peptide. No immunoreactivity was observed in this condition.
4
Immunophenotypic Analysis of Single Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single cells were collected with 0.25% trypsin-EDTA (Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA) and then resuspended in cold PBS at a concentration of 5×106 cells/mL. Then, the cells were incubated in the dark on ice for 30 min using antibodies including anti-CD29, anti-CD34, anti-CD45, anti-CD73, anti-CD90, anti-CD105 and anti-CD146. All antibodies were purchased from BioLegend, Inc. Subsequently, cells were twice washed and 500 μL PBS resuspension for each sample. The study was performed by Becton-Dickinson Accuri C6 (BD Biosciences, San Jose, CA, USA). FlowJo (version 10.0.7 r2) was used for the data analysis.
5
Immunohistochemical Profiling of Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical staining of the CD105, CD90, CD44 and Stro-1 molecules expression was performed using the following antibodies: anti-CD105 (mouse monoclonal, IgG2a, clone MEM-229; 1:300, Thermo Fisher, Scientific, Rockford, MI, USA), anti-CD90 (mouse monoclonal, IgG1, clone AF-9, 1:200, Thermo Fisher Scientific, Rockford, MI, USA), anti-CD44 (mouse monoclonal, IgG2a, clone 156-3C11, 1:4000, Thermo Fisher Scientific, Rockford, MI, USA), anti-STRO-1 (mouse monoclonal, IgM, STRO-1, 1:50, Abcam, Inc, Cambridge, UK), anti-osteocalcin (mouse monoclonal, IgG1, clone (OCG4), 1:400 (Thermo Fisher Scientific, Rockford, MI, USA), anti-osteopontin (mouse monoclonal, clone 7C5H17, 1:200, Abcam, Inc., Cambridge, UK), anti-bone sialoprotein (rabbit polyclonal, 1:50, Abcam, Inc., Cambridge, UK), anti-dentin sialophosphoprotein (rabbit polyclonal, 1:200, Abcam, Inc., Cambridge, UK).
6
Phenotypic Characterization of Cultured MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cultured MSCs at passage 5 were detached using 0.25% trypsin/EDTA and centrifuged at 400 × g for 5 min. The supernatant was discarded and the cell pellet was re-suspended in D-PBS containing 1% fetal bovine serum to the concentration of 105 cells/ml. Cells were then incubated with 10 µl of phycoerythrin-conjugated antibodies anti-CD34 (1:50, cat. no. MA1-19645; Thermo Fisher Scientific, Inc.), anti-CD44 (1:50, cat. no. MHCD4404; Thermo Fisher Scientific, Inc.), anti-CD73 (1:50, cat. no. FAB5795P; R&D Systems, Inc., Minneapolis, MN, USA), anti-CD90 (1:50, cat. no. A15794; Thermo Fisher Scientific, Inc.), anti-CD105 (1:50, cat. no. MA1-80944; Thermo Fisher Scientific, Inc.) at 25°C for 45 min. Then cells centrifuged at 300 × g for 15 min and washed with flow cytometry washing buffer (PBS with 0.5–1% bovine serum albumin; Sigma-Aldrich; Merck Millipore), the cells analyzed by flow cytometry analyzer (BD FACSARIA III, BD Biosciences).
7
Mesenchymal Stem Cell Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eight cell surface markers (CD105, CD73, CD90, CD45, CD34, CD14, CD19, and HLA-DR) were evaluated to identify MSCs according to ISCT criteria (Dominici et al., 2006 (link)). MSCs were counted to ensure each cell suspension had more than 1 × 106 cells. The cells were incubated with human anti-CD105 (PE), anti-CD73 (FITC), anti-CD90 (PE), anti-CD45 (PE), anti-CD34 (PE), anti-CD14 (FITC), anti-CD19 (APC), and anti-HLA-DR (FITC) (all from eBioscience, San Diego, CA, USA), respectively for 30 min at room temperature. The cells were then washed and suspended for flow cytometry analysis.
8
Comprehensive Antibody Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: anti-ALB (Abcam, ab8940, 1:400); anti-β-actin (Santa Cruz Biotechnology, sc-130301, 1:5 000); anti-caldesmon (Sigma-Aldrich, C4562, 1:300); anti-CD73 (BD Biosciences, 550741, 1:50); anti-CD90 (BD Biosciences, 555595, 1:100); anti-CD105 (eBioscience, 1-1057, 1:100); anti-CENPA (Abcam, ab13939, 1:400); anti-fibrillarin (Abcam, ab4566, 1:100); anti-FLAG (Sigma-Aldrich, M2, 1:2 000 for western blotting, 1:400 for immunofluorescence); anti-GAL4 (Abcam, ab14477, 1:1 000 for western blotting, 1:400 for immunofluorescence); anti-γH2AX (Millipore, 05-636, 1:400); anti-IgG-APC (eBioscience, 555751, 1:100); anti-IgG-FITC (eBioscience, 555748, 1:100); anti-IgG-PE (eBioscience, 555749, 1:100); anti-MAP2 (Sigma-Aldrich, m4403, 1:500); anti-NANOG (Abcam, ab21624, 1:250); anti-nestin (Millipore, MAB5326, 1:500); anti-NeuN (Millipore, ABN78 1:400); anti-OCT4 (Santa Cruz Biotechnology, sc-5279, 1:100); anti-Nucleolin (Abcam, ab22758, 1:200); anti-PAX6 (Covance, PRB-278P, 1:500); anti-SMA (Sigma-Aldrich, A5228, 1:100); anti-SM22 (Abcam, ab14106, 1:200); anti-SOX2 (Santa Cruz Biotechnology, sc-17320, 1:100); anti-Tuj1 (Sigma-Aldrich, T2200, 1:500); Alexa Fluor 555-conjugated wheat germ agglutinin (Thermo Fisher, W32464, 1:500). Other reagents, including deoxycytidine, hydroxyurea, NAC, nocodazole, and thymidine, were purchased from Sigma-Aldrich.
9
Multimarker Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For flow cytometry analysis, 2 × 105 cells were washed with phosphate-buffered saline (PBS) and then stained with the following antibodies: anti-CD11b (BD Pharmingen, Franklin Lakes, NJ, USA), anti-CD29 (eBioscience, San Diego, CA, USA), anti-CD31 (BD Pharmingen), anti-CD34 (BD Pharmingen), anti-CD44 (BD Pharmingen), anti-CD45 (BD Pharmingen), anti-CD73 (eBioscience), anti-CD105 (eBioscience), anti-CD90 (BD Pharmingen), anti-CD117 (BD Pharmingen), anti-Sca1 (BD Pharmingen), anti-CD26 (eBioscience), anti-EpCAM (Abcam), and goat anti-rabbit IgG PE-Cy5.5 (Thermo Fisher Scientific).
10
Phenotypic Characterization of BMMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell phenotypes of cultured BMMSCs at P2 were detected by flow cytometric analysis. For identification of the expression of mesenchymal stem cell surface markers, cells were harvested and washed by PBS. Then, the single‐cell suspension was incubated with fluorescein isothiocyanate (FITC)‐conjugated mouse anti‐CD11b, anti‐CD29, anti‐CD45, anti‐Sca‐1 and allophycocyanin (APC)‐conjugated anti‐CD34, anti‐CD90, anti‐CD105 and anti‐CD146 (all from eBioscience), respectively. Related conjugated IgG was used as control. Finally, cells were washed twice in PBS and subjected to flow cytometric analysis with Beckman Coulter CytoFLEX (Beckman Coulter).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!