Seven days after electroporation 5 × 105 T cells were collected and stained in 100 μL with fluorophore-conjugated anti-human antibodies for CD3 (BD Biosciences #564001), beta-2-microglobulin (BioLegend #316306) and Fixable Viability Dye eFluor780 or LIVE/DEAD Fixable Aqua Dead Cell Stain (ThermoFisher #65-0865-14 1:500 dilution, #L34966 1:40 dilution) were used to assess cell viability. Events were acquired on LSR II or LSRFortessa flow cytometers using FACSDiva software, and data was analyzed using FlowJo v10 software. See Supplementary Fig. 4 for gating strategy. The percent of positive events were normalized, by dividing over the sample percent positivity over pulse alone control percent positivity. Protein loss was then calculated as [1—% normalized sample positivity].
Fluorophore conjugated anti human antibodies for cd3
Manufactured by BD
Fluorophore-conjugated anti-human antibodies for CD3 are laboratory reagents used to detect and quantify CD3-positive cells in samples. These antibodies are conjugated with fluorescent dyes, allowing the labeled cells to be identified and analyzed using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
2 protocols using fluorophore conjugated anti human antibodies for cd3
1
Quantifying Protein Loss in T Cells
2
Quantifying T-cell Protein Loss
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Seven days after electroporation a total of 5 x 106 T cells were collected and stained in 100 uL with fluorophore-conjugated anti-human antibodies for CD3 (BD Biosciences #564001), beta-2microglobulin (BioLegend #316306) and Fixable Viability Dye eFluor780 or LIVE/DEAD Fixable Aqua Dead Cell Stain (ThermoFisher #65-0865-14 1:500 dilution, #L34966 1:40 dilution) were used to assess cell viability. Events were acquired on LSR II or LSRFortessa flow cytometers using FACSDiva software, and data was analyzed using FlowJo v10 software. See Supplementary Fig. 4 for gating strategy. The percent of positive events were normalized, by dividing over the percent positivity seen in pulse alone controls. Protein loss was then calculated as [1 -% normalized sample positivity].
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