Cell surface markers were measured on 50%, 80% and 100% confluent BMSCs by Flow Cytometry assay; the markers were selected according to either global gene expression profiling results or from the literature34 (link)–36 (link). The following antibodies were used, PODXL-FITC (MBL International Inc, Nagoya, Japan), CD49f-PE, CD54-APC, TLR1-PE, CD106-PE, LAMP1 (CD107a)-APC, CD146-PE, CCR7-PE, CD55-FITC, TLR4-PE (BD Biosciences), CD200-APC, CD10-APC (Biolegend, San Diego, CA, USA), and FZD4-APC (R&D, Minneapolis, MN, USA), isotype controls were IgG1-FITC, IgG1-PE, IgG2a-FITC (BD Bioscience), IgG1-APC (eBioscience, San Diego, CA, USA). In brief, BMSCs were incubated with antibody cocktail for 30 minutes in the dark at room temperature and washed with PBS containing 1% BSA; the cells were then suspended in 0.3 mL PBS with 1% BSA, counterstained with 7-AAD; twenty-thousand events were acquired on an Accuri C6 flow cytometer. The data were analyzed with FlowJo software (TreeStar, Ashland, OR). Positive cells were identified as those whose intensities were greater than 99 percentile of the isotype control.
Apc igg1
Manufactured by Thermo Fisher Scientific
Sourced in United States
The APC-IgG1 is a fluorescently labeled antibody used for flow cytometry applications. It is conjugated with the fluorescent dye Allophycocyanin (APC), which allows for the detection of target cells or proteins.
Automatically generated - may contain errors
Lab products found in correlation
Cd49f pe, by BD (1 mentions)
Cd146 pe, by BD (1 mentions)
Igg1 pe, by BD (1 mentions)
Igg1 fitc, by BD (1 mentions)
Ccr7 pe, by BD (1 mentions)
Igg2a fitc, by BD (1 mentions)
Cd10 apc, by BioLegend (1 mentions)
Cd54 apc, by BD (1 mentions)
Lamp1, by BD (1 mentions)
Tlr4 pe, by BD (1 mentions)
Cd106 pe, by BD (1 mentions)
Cd107a apc, by BD (1 mentions)
Igg1 fitc, by Agilent Technologies (1 mentions)
Igg1 pe, by Agilent Technologies (1 mentions)
Facscalibur instrument, by BD (1 mentions)
Anti cd3 fitc, by Agilent Technologies (1 mentions)
Apc anti nkg2d, by Thermo Fisher Scientific (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti foxo1, by Cell Signaling Technology (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
5 protocols using apc igg1
1
Flow Cytometric Analysis of BMSC Markers
2
Immunophenotyping of NK Cells
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For staining of peripheral blood mononuclear cells with purified antibodies, 100,000 cells were incubated with 0.2μg of antibody diluted in PBS/0.5% BSA/0.02% NaN3 (FACS medium) for 1h on ice as previously described [42 (link), 43 (link)]. Cells were centrifuged 400g/5min and supernatant was removed. The cells were incubated 30mins on ice with secondary antibodies, washed with FACS medium and analyzed with FACS calibur instrument (BD, Franklin Lakes, NJ, USA) and FlowJo or CellQuest software. The following antibodies were used: PE-anti-CD56 (DAKO, Glostrup, Denmark, clone MOC-1), FITC-anti CD3 (DAKO, clone UCHT1), APC-anti-NKG2D (eBioscience, San Diego, USA, clone CX5), APC-anti-NKp46 (eBioscience). Isotype control IgG1-FITC (DAKO), IgG1-PE (DAKO) and IgG1-APC (eBioscience) served as negative controls. NK cells were gated based on positive CD56 expression and lack of CD3 expression. At least 2,500 gated NK cells were analyzed in each reading.
3
Integrin Expression in SKOV3 and OVCAR-3 Cells
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The cultured monolayer of SKOV3 and OVCAR-3 cells and MCAs were harvested. The different groups of cells were stained in duplicate with FITC-anti-α3β1, APC-anti-α3 and PE-α4 or control FITC-IgG1, APC-IgG1 or PE-IgG1 (eBioscience). In addition, some cells were stained with FITC-anti-β1 and PE-anti-α5 or control IgG. After being washed, the cells were analyzed by flow cytometry.
4
Immunophenotyping of B-cell Subsets
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Cells were surface-stained and analyzed by flow cytometry on LSRII flow cytometers (Becton Dickinson) using Cy5-, APC- or FITC-anti-IgM (μ-chain-specific, Southern Biotech), PE-anti-λ5 (LM34, a gift from A. G. Rolink), PE-Cy7-B220 (eBioscience), PE-B220 (BD Pharmingen) APC-CD19 (eBioscience), PE-CD43 (BD Pharmingen), PE-κ (Southern Biotech), PE-CD11b (eBioscience) and APC-CD127 (eBioscience). Intracellular stainings were performed using Fix and Perm cell permeabilization Kit (ADG) according to the manufacturer’s instructions. Antibodies used for intracellular FACS-staining were PE-anti-Pax5 (eBioscience), anti-SLP-65 (monoclonal IgG mouse, self-produced), anti-AKT (Cell Signaling), anti-FoxO1 (Cell Signaling), pAKT (Cell Signaling), APC-anti-rabbit IgG1 (AbD Serotec) and APC-anti-mouse IgG (AbD Serotec). PE-IgG2a κ (eBioscience), APC-IgG1 (eBioscience), APC-IgG2a κ (eBioscience) were used as isotype controls. PE-anti-Biotin (BioLegend), Alexa Fluor 647-anti-mouse-IgG Fab2 and Alexa Fluor 647-anti-rabbit-IgG Fab2 were used as secondary antibodies or as staining controls, respectively when used without the primary antibody.
5
Endothelial Differentiation and VE-Cadherin Analysis
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After 7 days of endothelial differentiation, the infected DPSCs or HUVECs were trypsinized, followed by resuspension in PBS. Next, the cells were incubated for 30 min at 4 °C with APC-conjugated monoclonal antibody against VE-Cadherin (17-1449-42, eBioscience). The isotypic control used was APC-IgG1 (17-4714-81, eBioscience). The analysis of the stained cells was performed with a flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA).
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