The following antibodies were used for these studies: CD4-Alexa Fluor 700 (catalog #56-0048-82), CD5-PE-Cy7 (#25-0059-42), CD7-Alexa Fluor 700 (#56-0079-42), CD31-PerCP/eFluor710 (#46-0319-42), CD45-APC-eFluor780 (#47-0459-42), and CD144-PE (#12-1449-82) from eBioScience; CD8-PE (#555367), CD34-PE (#550761), and CD34-APC (#555824) from BD Pharmingen; KDR-PE (#359904) from BioLegend; and KDR-PE (#FAB357P100) from R&D Systems. Stained cells were analyzed using an LSRII (BD Biosciences) flow cytometer at the indicated time points. Data analysis was performed using FlowJo software. For T-lymphoid studies, analyses were carried out by gating on live cells, as indicated by lack of DAPI uptake, followed by gating on CD45+ cells. Gates were set using appropriate isotype controls.
Cd144 pe
Manufactured by Thermo Fisher Scientific
Sourced in Germany
CD144-PE is a fluorescently-labeled antibody that binds to the CD144 (also known as VE-cadherin) cell surface antigen. CD144 is a marker for endothelial cells and is commonly used in flow cytometry applications to identify and characterize this cell population.
Automatically generated - may contain errors
Lab products found in correlation
Cd5 pe cy7, by Thermo Fisher Scientific (2 mentions)
Cd184 brilliant violet 421, by BioLegend (2 mentions)
Cd34 pe cy7, by Thermo Fisher Scientific (2 mentions)
Facsaria 2, by BD (2 mentions)
Cd34 pe, by BD (1 mentions)
Cd45 apc efluor780, by Thermo Fisher Scientific (1 mentions)
Cd4 alexa fluor 700, by Thermo Fisher Scientific (1 mentions)
Cd8 pe, by Thermo Fisher Scientific (1 mentions)
Cd34 apc, by BD (1 mentions)
Cd7 alexa fluor 700, by Thermo Fisher Scientific (1 mentions)
Kdr pe, by R&D Systems (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Cd40 apc, by Miltenyi Biotec (1 mentions)
Il 13, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
C kit apc, by Thermo Fisher Scientific (1 mentions)
Cd34 apc, by Thermo Fisher Scientific (1 mentions)
Cd43 pe, by Thermo Fisher Scientific (1 mentions)
Cd4 pe, by Thermo Fisher Scientific (1 mentions)
5 protocols using cd144 pe
1
Multicolor Flow Cytometry Panel for Immune Cell Profiling
2
In Vitro Macrophage and HUVEC Priming
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For simulating inflammatory conditions in vitro, macrophages were primed with fresh medium supplemented with 20 ng/ml IFNγ (Gibco, Thermo Fisher Scientific) and/or 100 ng/ml lipopolysaccharides (LPS) (Sigma) to generate pro-inflammatory M1 macrophages. For anti-inflammatory conditions, macrophages were primed with 20 ng/ml IL-4 (PeproTech, London, UK) and/or 20 ng/ml IL-13 (PeproTech) to generate anti-inflammatory M2 macrophages. The incubation time was 24 h for all the conditions. PKH-MP were then added to macrophages (ratio 1:50,000) and incubated for another 24 h after which uptake was analyzed by flow cytometer and confocal microscopy.
HUVEC were primed with fresh medium supplemented with 10 ng/ml TNFα and/or 50 ng/ml IFNγ for 24 h. MP were added to the cells (ratio 1:50,000) and incubated for another 24 h after which uptake was analyzed by flow cytometer. After 48 h, HUVEC were collected for functional analysis by flow cytometer, after staining with HLA-I-PacBlue (BD Biosciences, San Jose, CA), HLA-II-PerCP (BioLegend), CD40-APC (Miltenyi Biotec, Bergisch Gladbach, Germany), CD144-PE (Ebioscience, Thermo Fisher Scientific).
HUVEC were primed with fresh medium supplemented with 10 ng/ml TNFα and/or 50 ng/ml IFNγ for 24 h. MP were added to the cells (ratio 1:50,000) and incubated for another 24 h after which uptake was analyzed by flow cytometer. After 48 h, HUVEC were collected for functional analysis by flow cytometer, after staining with HLA-I-PacBlue (BD Biosciences, San Jose, CA), HLA-II-PerCP (BioLegend), CD40-APC (Miltenyi Biotec, Bergisch Gladbach, Germany), CD144-PE (Ebioscience, Thermo Fisher Scientific).
3
Multicolor Flow Cytometry Immunophenotyping
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The following antibodies were used for these studies: CD34-APC (clone 8G12, 1:100), CD34-PE-CY7 (clone 4H11, 1:100), CD43-PE or FITC (clone 1G10, 1:30 and 1:10 respectively), CD4-PE-Cy7 (clone RPA-T4, 1:100), CD8-PE (clone RPA-T8, 1:30), CD45-APC-Cy7 or eFluor450 (clone 2D1, 1:50), CD56-APC (clone B159, 1:30), CD144-PE (clone 123413, 1:50), CD184-Brilliant Violet 421 (clone 12G5, 2:100), CD73-APC (clone AD2, 1:400), KDR (clone 89106, 1:7) and CD235a-APC (clone HIR2, 1:50). Most antibodies were purchased from BD Biosciences (San Diego, CA). The exceptions are CD34-PE-CY7 and CD144-PE that were purchased from eBioscience, CD184-Brilliant Violet 421 purchased from Biolegend and KDR purchased from R&D systems. Cells were sorted with FACSAria™II (BD) cell sorter at the Sick Kids/UHN Flow Cytometry Facility.
4
Multicolor Flow Cytometry Immunophenotyping
5
Evaluating hPSC-Derived Cells by Flow Cytometry
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To evaluate HE differentiation by flow cytometric analysis, hPSC-derived cells were dissociated by incubation in 0.1% collagenase for 2 h followed by TypLE Express for 8 min. Cells were mechanically dissociated and resuspended in 2% v/v FBS in Hank's buffered saline solution (HF). The following antibodies (obtained from BD Biosciences unless otherwise specified) were used at indicated dilutions: CD34-APC (1:100); CD144-PE (4:100); CD144-V450 (4:100); CD43-PE (3:100); CD45-PE-CY7 (4:100); CD4-PE (1:50) or CD34-PE (1:50); CD5-PE/Cy7 (1:200); CD7-AF700 (1:200); CD43-APC (1:200); CD45-APC/eF780 (1:200, eBiosciences); Ckit-APC (1:100); and PE-KDR (1:50). Control samples were prepared by incubating duplicate cell samples with respective fluorochome-labelled isotype antibodies. Samples were incubated with antibodies for 35 min on ice and then washed three times in cold HF. Viability discrimination was performed simultaneously using the viability stain 7-amino-actinomycin D at 1 μl ml−1 (Molecular Probes). Samples were analysed on a Becton Dickinson FACS Fortessa machine using BD FACS Diva Software. Cells were sorted using a FACS Aria (BD Bioscience) cell sorter (Donnelly Centre for Cellular & Biomolecular Research).
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