Flow cytometry analysis was performed on a Guava easyCyte 6HT 2L flow cytometer (Merck Millipore, Darmstadt, Germany). Briefly, cells were harvested by trypsin at 37°C for 5 minutes, centrifuged at 400*g and resuspended in 1X PBS supplemented with 1% FCS. 4000 cells were incubated with 10 μL anti-SSEA4-PE (BD Biosciences, Pharmingen™, San Diego, USA), 5 μL IgG1-PE (Beckman Coulter Inc., France), 5 μL anti-CD56-PE (Beckman Coulter Inc., France) and 10 μL anti-CD90-PE (Beckman Coulter, Inc., France) for 15 minutes in a 1.5 mL Eppendorf tube at 4°C in dark. Cell were washed with 1 mL PBS, centrifuged at 400*g and resuspended in 200 μL of 1X PBS in a 96- well round bottom plate. After washing and resuspension, each reaction received 5 μL of viability dye 7-aminoactinomycin D (Beckman Coulter Inc., France) and plate was incubated for 10 minutes at 4°C. Cell events were acquired with Guava InCyte™ v.2.3 software. Histograms and dot-plots were generated with a minimum of 3000 events with a sample flow rate of 1.8 μL/mL. Positive staining was obtained by comparison with isotype control set as 99% negative.
Igg1 pe
Manufactured by Beckman Coulter
Sourced in United States
IgG1-PE is a fluorescently-labeled antibody used in flow cytometry applications. It is a mouse IgG1 isotype conjugated to the fluorescent dye Phycoerythrin (PE). IgG1-PE can be used to detect and quantify target cells expressing the relevant antigen.
Automatically generated - may contain errors
Lab products found in correlation
Igg1 fitc, by Beckman Coulter (4 mentions)
Guava easy cyte 6ht 2l flow cytometer, by Merck Group (2 mentions)
Cd34 pe, by Beckman Coulter (2 mentions)
Cd105 pe, by Beckman Coulter (2 mentions)
Anti cd56 pe, by Beckman Coulter (1 mentions)
Anti ssea4 pe, by BD (1 mentions)
Anti cd90 pe, by Beckman Coulter (1 mentions)
7 aminoactinomycin d, by Beckman Coulter (1 mentions)
Accuri flow cytometer, by BD (1 mentions)
Fitc conjugated anti cd90, by BioLegend (1 mentions)
Pe anti cd59, by BioLegend (1 mentions)
Cytomics fc500 flow cytometer, by Beckman Coulter (1 mentions)
Mouse igg1 fitc, by Beckman Coulter (1 mentions)
Cd56 pe, by Beckman Coulter (1 mentions)
Cd19 pe, by BioLegend (1 mentions)
Cd73 pe, by BD (1 mentions)
Cd90 pe, by Beckman Coulter (1 mentions)
Cd45 pe, by Beckman Coulter (1 mentions)
Cd14 pe, by Thermo Fisher Scientific (1 mentions)
Hla dr pe, by Beckman Coulter (1 mentions)
6 protocols using igg1 pe
1
Flow Cytometry Analysis of Stem Cell Markers
2
Quantifying Surface Markers in Human Skin Fibroblasts
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For measurement of CD90, CD13, CD55 and CD59 surface expression, HSF were detached from 6-well plates with EDTA 5mM, washed with PBS/BSA, and incubated for one hour with the following monoclonal antibodies: phycoerythrin (PE)-conjugated anti-CD55 (1:100, BioLegend), Fluorescein isothiocyanate (FITC) conjugated anti-CD90 (1:100, BioLegend), PE anti-CD59 (1:100, BioLegend), PE anti-CD13 (1:100, BECKMAN COULTER) or isotype control antibodies: IgG1-FITC (1:100, BECKMAN COULTER) and IgG1-PE (1:100, BECKMAN COULTER). Stainings were visualized by flow cytometry with BD ACCURI flow cytometer.
3
Characterization of Hematopoietic Stem and Progenitor Cells
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Purified CD34+ cells have been reported to be enriched in HSPCs [9 (link)]. The enrichment of HSPCs in CD34+ cells has previously been investigated by profiling cell surface antigens using flow cytometry. In the present study, the CD34+ cells in CB were incubated for 30 min at 4°C with FITC-conjugated anti-human CD34 (Beckman Coulter Immunotech, Marseille, France) and each of the following fluorescent monoclonal antibodies: PE-conjugated anti-human CD38, PC5-conjugated anti-human CD45, PE-conjugated anti-human CD41, PC5-conjugated anti-human CD45RA (Beckman Coulter, Brea, USA), PE-conjugated anti-human Tie-2 (R&D Systems, Minneapolis, MN), PE-conjugated anti-human CD110, PC5-conjugated anti-human CD117 and PC5-conjugated anti-human CD123 (Becton Dickinson Biosciences, San Jose, CA). After incubation, the CD34+ cells were washed and analyzed using a Cytomics FC500 flow cytometer (Beckman Coulter Immunotech, Marseille, France). A combination of isotype-matched monoclonal antibodies, mouse IgG1-FITC, IgG1-PE and IgG1-PC5 (Beckman Coulter Immunotech, Marseille, France) was used as a negative control.
4
Multicolor Flow Cytometry Analysis
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Flow cytometry analysis was performed on a Guava easyCyte 6HT 2 L flow cytometer (Merck Millipore, Darmstadt, Germany) as described before [29 (link)]. Before measurement, 40,000 cells were resuspended in 195 μl 1× PBS and incubated after addition of 5 μl CD34-PE, CD56-PE, CD146-PE, IgG1-PE, IgG1-FITC, CD90-PE, CD105-PE, HLA-DR-PE, CD45-PE (all from Beckman Coulter, CA, USA), CD49a-FITC (MiltenyiBiotec GmbH, Bergisch Gladbach, Germany), CD14-PE (Thermo Scientific, MA, USA), CD19-PE (BioLegend, CA, USA), or CD73-PE (Becton Dickinson, NJ, USA) for 30 min at 4 °C in the dark following a washing step with 1× PBS. Histograms and dot plots were generated with a minimum of 5000 events at a sample flow rate of 1.8 μl/ml. Positive staining was obtained by comparison with isotype control set as 99% negative or comparison to control (negative) cells.
5
NK Cell Immunophenotyping with Flow Cytometry
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NK cells were incubated with anti-human CD3-FITC (1:100; Beckman Coulter, Inc.; cat. no. A07746), anti-human CD16-PE (1:100; Beckman Coulter, Inc.; cat. no. A07766), anti-human CD56-PE-cyanine5 (1:100; Beckman Coulter, Inc.; cat. no. A07789), anti-human CD314-PE (1:100; Beckman Coulter, Inc.; cat. no. A08934), anti-human CD226 [DNAX accessory molecule (DNAM)-1]-FITC (1:50; BD Pharmigen; BD Biosciences; cat. no. 559788), or anti-human CD244 (2B4)-FITC (1:50; BD Pharmigen; BD Biosciences; cat. no. 550815) for 20 min in the dark at room temperature. The cells were also incubated with each isotype control: IgG1-FITC (1:100; Beckman Coulter, Inc.; cat. no. A07795), IgG1-PE (1:100; Beckman Coulter, Inc.; cat. no. A07796), IgG1-PE-cyanine5 (1:100; Beckman Coulter, Inc.; cat. no. A07798), IgG1-FITC (1:50; BD Pharmigen; BD Biosciences; cat. no. 555748), and IgG2a-FITC (1:50; BD Pharmigen; BD Biosciences; cat. no. 555573) for 20 min in the dark at room temperature and evaluated by FCM.
6
Comparative Analysis of hBM-MSCs and hWJ-MSCs Phenotypes
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hBM-MSCs and hWJ-MSCs were detached and counted; 1 × 105 cells were incubated at RT for 20 min with the following directly conjugated mouse-anti human antibodies: CD90 FITC (Beckman Coulter, Fullerton, CA, USA), CD73 APC (Miltenyi Biotec, Gladbach, Germany), CD105 PE (Beckman Coulter, Fullerton, CA, USA), CD45 PC7 (Beckman Coulter, Fullerton, CA, USA), HLA class-II FITC (Beckman Coulter, Fullerton, CA, USA), CD14 PC7 (Beckman Coulter, Fullerton, CA, USA), and CD34 PE (Beckman Coulter, Fullerton, CA, USA). The isotype-matched immunoglobulins IgG1 FITC (Beckman Coulter, Fullerton, CA, USA), IgG1 PE (Beckman Coulter, Fullerton, CA, USA), IgG1 APC (Beckman Coulter, Fullerton, CA, USA), and IgG1 PC7 (Beckman Coulter, Fullerton, CA, USA) were used as negative controls under the same conditions. At least 15,000 total events were acquired with a BD FACSVerse flow cytometer (Becton Dickinson, BD, NJ, USA). Further analysis and plots were generated using the BD FACSuite analysis software. Statistics are summarized in Table S1 (Supplementary Materials) .
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