Cultures of Jurkat E6-1 (ATCC) and lentivirus transducted Jurkat E6-1 cells that express Zaire Ebolavirus glycoprotein on the surface (gift from C. Davis and R. Ahmed, Emory University School of Medicine) were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS, HyClone) according to ATCC recommendations. Cells were washed with ice-cold FACS buffer (Dulbecco's PBS containing 2% FBS and 50 nM Dasatinib), counted, seeded at ∼50,000 viable cells per well in V-bottom 96-well plate for each mAb to be tested and incubated 60 min at 4 °C with serial tenfold dilutions of mAb in a total volume 100 μl per staining. Cells were washed with FACS buffer by centrifugation for 2 min at 800g followed by incubation with 1:500 dilution of secondary goat anti-human IgG PE Ab (SouthernBiotech) in FACS buffer. After washing, 5,000–10,000 live cell events were acquired using a three-laser LSR-II flow cytometer (BD Biosciences) and analysed with FlowJo software (Tree Star). The dead cell population was excluded using propidium iodide staining.
Goat anti human igg pe ab
Manufactured by Southern Biotech
Goat anti-human IgG PE Ab is a laboratory reagent that binds to human immunoglobulin G (IgG) antibodies. The PE (phycoerythrin) fluorescent dye is conjugated to the goat anti-human IgG antibody, allowing for the detection and quantification of human IgG in samples using flow cytometry or other fluorescence-based techniques.
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2 protocols using goat anti human igg pe ab
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Jurkat E6-1 Cells Expressing Zaire Ebolavirus Glycoprotein
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Jurkat E6-1 Cells Expressing Zaire Ebolavirus Glycoprotein
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Cultures of Jurkat E6-1 (ATCC) and lentivirus transducted Jurkat E6-1 cells that express Zaire Ebolavirus glycoprotein on the surface (gift from C. Davis and R. Ahmed, Emory University School of Medicine) were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS, HyClone) according to ATCC recommendations. Cells were washed with ice-cold FACS buffer (Dulbecco's PBS containing 2% FBS and 50 nM Dasatinib), counted, seeded at ∼50,000 viable cells per well in V-bottom 96-well plate for each mAb to be tested and incubated 60 min at 4 °C with serial tenfold dilutions of mAb in a total volume 100 μl per staining. Cells were washed with FACS buffer by centrifugation for 2 min at 800g followed by incubation with 1:500 dilution of secondary goat anti-human IgG PE Ab (SouthernBiotech) in FACS buffer. After washing, 5,000–10,000 live cell events were acquired using a three-laser LSR-II flow cytometer (BD Biosciences) and analysed with FlowJo software (Tree Star). The dead cell population was excluded using propidium iodide staining.
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