CAOV3, SKOV3, and HEY1 lines were purchased from ATCC. A2780 was provided by Dr. S. Murphy (Duke University), Kuramuchi was provided by Dr. Deborah Marsh (University of Sydney). OVCAR3 was proved by Dr. Kathleen Cho (University of Michigan). Ovarian cancer cell lines were cultured in Roswell Park Memorial Institute (RPMI)-1640 Media, supplemented with 10% fetal bovine serum (FBS), as previously described [13 (link)]. Anti-CD105 antibodies were purchased from Abcam (APC-conjugated, for FACS analysis) and eBioscience (SN6, for in vitro cell blocking). Smad2/3 and phospho-Smad2/3 antibodies were purchased from Cell Signaling Technology; GAPDH was purchased from Proteintech.
Anti cd105 antibody
Manufactured by Abcam
Sourced in United Kingdom
Anti-CD105 antibody is a laboratory reagent used for the detection and analysis of CD105, also known as endoglin, a cell surface protein. This antibody can be employed in various immunological techniques, such as flow cytometry, immunohistochemistry, and Western blotting, to study the expression and distribution of CD105 in biological samples.
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Lab products found in correlation
Caov3, by American Type Culture Collection (1 mentions)
Skov3, by American Type Culture Collection (1 mentions)
Gapdh, by Proteintech (1 mentions)
Anti cd73, by BD (1 mentions)
Alcian blue, by Merck Group (1 mentions)
Phycoerythrin conjugated anti cd34, by Abcam (1 mentions)
Anti cd49d, by BD (1 mentions)
Cytoflex s flow cytometer, by Beckman Coulter (1 mentions)
Anti stro 1, by R&D Systems (1 mentions)
Anti lgr5, by Abcam (1 mentions)
Anti sox2, by R&D Systems (1 mentions)
Anti lsd1, by Merck Group (1 mentions)
Anti sox9, by Abnova (1 mentions)
3 3 diaminobenzidine (dab), by Abcam (1 mentions)
Anti gli1, by Abcam (1 mentions)
Anti cd44, by Abcam (1 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
Anti ssea3 antibody, by Abcam (1 mentions)
Anti cd24 antibody, by Abcam (1 mentions)
5 protocols using anti cd105 antibody
1
Ovarian Cancer Cell Culture and Antibodies
2
Phenotypic Characterization of ASCs
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ASCs from passage 3 were incubated with Fluorescein isothiocyanate-conjugated anti-cluster of differentiation (CD)90 antibodies (1/20, Abcam, Cambridge, United Kingdom) and anti-CD105 antibodies (1/20, Abcam) or with Phycoerythrin-conjugated anti-CD34 (1/20, Abcam), anti-CD49d (1/20, Becton, Dickinson and Company), and anti-CD73 (1/20, Becton, Dickinson and Company) antibodies for 30 minutes at room temperature. As a control, cells were stained with an isotype control Immunoglobulin G (IgG). Cells were subsequently washed with phosphate buffer saline (PBS) and analyzed on CytoFLEX S flow cytometer (Beckman Coulter, Chino, California, USA) using CytExpert software.
ASCs were then differentiated into adipogenic or chondrogenic cells using adipogenesis or chondrogenesis differentiation kits (GIBCO). The in vitro differentiation potential was determined by lineage-specific staining; adipogenic ASCs were stained with Oil Red O and chondrogenic ASCs were stained with Alcian Blue (Sigma, St. Louis, Missouri, USA).
ASCs were then differentiated into adipogenic or chondrogenic cells using adipogenesis or chondrogenesis differentiation kits (GIBCO). The in vitro differentiation potential was determined by lineage-specific staining; adipogenic ASCs were stained with Oil Red O and chondrogenic ASCs were stained with Alcian Blue (Sigma, St. Louis, Missouri, USA).
3
Evaluating Stem Cell Markers in MCL Repair
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Four days after transection, both the right and left MCL of 4 rats treated with 0.5 μg/μl rHAM+ dissolved in PGA carrier, and 5 rats treated only with PGA carrier, were harvested. The MCLs were fixed in 4% para‐formaldehyde for 24 hrs at 4°C, and incubated in 30% sucrose solution at 4°C until the tissue soaked in the tube. The tissues were then embedded in O.C.T compound (Sakura Fintek Inc, Torrance, CA, USA), frizzed to −80°C and cryo‐sectioned (Leica Biosystems, Wetzlar, Germany; 8 μm thick). Indirect immunohistochemistry was performed using anti CD105 antibody (Abcam plc, Cambridge, UK), and immuno‐fluorescence using anti STRO‐1 (R&D Systems Inc., Minneapolis, MI, USA) and anti Ki67 antibody (Bioscience Inc., San Diego, CA, USA).
4
Immunohistochemical Analysis of Stem Cell Markers
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After routinely dewaxing and hydration, sections proceed to be antigen repaired with TE buffer at 98 °C. Each section was blocked with 3% H2O2. Each section was incubated with anti-Gli1 (Abcam), anti-CD44 (Abcam), anti-LSD1 (Sigma), anti-Sox2 (R&D), anti-Sox9 (Abnova), anti-LGR5 (Abcam), in primary antibody dilution buffer for 1 h at ambient temperature (AT). Then anti-mouse/rabbit antibody were used to incubated with tissue samples for 30 min at AT. Lastly, chromogenic agent 3, 3′-diaminobenzidine (Dako) was used to stain tissue samples.
The double immunostaining procedure was executed in the same section, the first step was to use anti-Gli1 antibody staining with 3,3′-diaminobenzidine, the second step was to use anti-CD105 antibody (Abcam) staining with AEC.
Two pathologists (WB Qi & YH Xuan) assessed the immunohistochemical results and the staining results were assessed according to previous study [9 (link)].
The double immunostaining procedure was executed in the same section, the first step was to use anti-Gli1 antibody staining with 3,3′-diaminobenzidine, the second step was to use anti-CD105 antibody (Abcam) staining with AEC.
Two pathologists (WB Qi & YH Xuan) assessed the immunohistochemical results and the staining results were assessed according to previous study [9 (link)].
5
Flow Cytometric Profiling of iPSC Markers
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Cell surface antigens for human iPSCs were analyzed with FACS. Cells (1 × 105 cells per well) were incubated with one of the following primary antibodies: anti-CD24 antibody (Abcam), 1:100 dilution; anti-SSEA3 antibody (Abcam), 1:100 dilution; anti-CD105 antibody (Abcam), 1:200 dilution; anti-Nanog antibody (CST), 1:100 dilution; anti-Sox2 antibody (CST), 1:300 dilution; and anti-Oct4 antibody, 1:600 dilution (CST). After 1 h of incubation at 4 °C, the cells were washed, centrifuged, stained with 100 μl of a secondary antibody selected from anti-mouse IgG (H + L) Alexa Fluor® 488 Conjugate (CST) at 1:1000 dilution, anti-rat IgG (H + L) Alexa Fluor® 488 Conjugate (CST) at 1:1000 dilution, and anti-rabbit IgG (H + L) Alexa Fluor® 488 Conjugate (CST) at 1:1000 dilution. The stained cell pellets were suspended in 200 μl PBS/10 % FCS at 4 °C for flow cytometry analysis. All antibodies were validated for antigen specificity.
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