The following antibodies directed against mouse antigens were used: Percp-CD45 (Biolegend 103130), FITC-CD31 (BD 553372), APC-CD117/Kit (BD 553356), PE-CD31 (Biolegend 102407), FITC-Ter119 (BD 561032), Percp-IgG2bk (Biolegend 400336), FITC-IgG2ak (BD 553929), APC-IgG2bk (BD 556924), PE-IgG (Biolegend 405307), PE-Annexin V (BD560930), DAPI (ThermoFisher D1306), PE-ItgαV (Biolegend 104106), and PE-Itgα4 (BD 557420). For flow analysis, antibodies were used at a concentration of 2 µg ml−1.
Pe igg
Manufactured by BioLegend
Sourced in United States
PE-IgG is a fluorochrome-conjugated antibody that can be used as an isotype control in flow cytometry experiments. It binds to immunoglobulin G (IgG) proteins and emits a fluorescent signal when excited by the appropriate wavelength of light.
Automatically generated - may contain errors
Lab products found in correlation
Cd31 fitc, by BD (1 mentions)
Annexin 5 pe, by BD (1 mentions)
Cd31 pe, by BioLegend (1 mentions)
Cd45 percp, by BioLegend (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Facsjazz, by BD (1 mentions)
Fitc igg, by BioLegend (1 mentions)
Accuri c6, by BD (1 mentions)
Rnase a, by Thermo Fisher Scientific (1 mentions)
Cd146, by BioLegend (1 mentions)
Accutase, by Innovative Cell Technologies (1 mentions)
4 protocols using pe igg
1
Multiparametric Flow Cytometry Analysis
2
Analyzing Tumor-Infiltrating Dendritic Cells
3
Flow Cytometry Analysis of Stem Cell Markers
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After treatment, the cells were digested and washed twice with cold PBS containing 2% BSA, then resuspended in 100 μL PBS and incubated with the antibody FITC-IgG, PE-IgG and FITC-CD44, PE-CD24 (BioLegend, San Diego, CA, USA) for 30 min at 4 °C in the dark. The expressions of stemness-related markers of CD24 and CD44 were detected by flow cytometry, and the ratio of CD44+ and CD24− cells was analyzed.
4
Stem Cell Marker Expression and Cell Cycle Analysis
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The cells (0.5 × 105/well) were seeded in 100 mm culture dishes and allowed to attach for 24 h. The cells were treated with the indicated drugs for 5 days and harvested after Accutase (Innovative Cell Technologies, CA, USA). Markers included antibodies targeting CD73 (Bio Legend, 344003, San Diego, CA, USA) and CD146 (Bio Legend, 361005) as positive stem cell markers. As controls, the cells were incubated with mouse isotype controls (PE-IgG, Bio Legend, 409303). Fluorescence of the stained cells was captured in an Accuri C6 (BD Biosciences, CA, USA), and after the acquisition of 100,000 events, the data were analyzed using FCS Express software. The data are expressed as the percentage of cells positive for each marker.
For cell cycle analysis, collected cells were washed with PBS and then fixed with ethanol at 4 °C for 30 min. Fixed cells were washed with PBS two times and incubated with RNase A (10 mg/mL, Thermo Scientific, EN0531) at room temperature for 5 min. The cells were stained with propidium iodide (1 mg/mL, Thermo Scientific, P3566, Waltham, MA, USA) for 5 min in the dark. Cell cycle analysis was performed using an Accuri C6 (BD Biosciences, CA, USA).
For cell cycle analysis, collected cells were washed with PBS and then fixed with ethanol at 4 °C for 30 min. Fixed cells were washed with PBS two times and incubated with RNase A (10 mg/mL, Thermo Scientific, EN0531) at room temperature for 5 min. The cells were stained with propidium iodide (1 mg/mL, Thermo Scientific, P3566, Waltham, MA, USA) for 5 min in the dark. Cell cycle analysis was performed using an Accuri C6 (BD Biosciences, CA, USA).
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