A volume of 200 µL blood was labeled with 20 µL fluorescence-conjugated antihuman antibodies (double staining with CD3 FITC/CD4 PE, CD3 FITC/CD8 PE, CD3 FITC/CD19 PE, CD3 FITC/CD16/CD56 PE, BD Pharmingen, San Diego, CA, USA) for 30 minutes following the manufacturer’s instructions and recommendations. Then the red blood cells were lysed, washed 3 times in phosphate-buffered saline (PBS), and then fixed in 4% paraformaldehyde in PBS, and cytometry was performed by using Cell Quest software on a FACScan flow cytometer (Becton Dickinson, Mountain View, CA, USA). Suitable negative isotype controls were used to rule out the background fluorescence. The data were generated by cytofluorometric analyses of 10,000 events. The positive population for each marker was determined using quadrant statistics. The data were analyzed using FlowJo 7.6 software.
Cd3 fitc cd19 pe
Manufactured by BD
Sourced in United States
The CD3 FITC/CD19 PE is a laboratory reagent used for the identification and enumeration of T cells (CD3+) and B cells (CD19+) in biological samples using flow cytometry. It contains fluorescently-labeled antibodies that bind to specific cell surface markers, allowing for the detection and quantification of these cell populations.
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2 protocols using cd3 fitc cd19 pe
1
Immunophenotyping by Flow Cytometry
2
Multicolor Flow Cytometry Analysis
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A sample of EDTA anticoagulated whole blood was incubated with BD Tritest CD4FITC/CD8PE/CD3PerCP reagent, another with CD3FITC/CD19PE, and the third one with CD45FITC/CD14PE (all reagents of BD Biosciences) and isotype controls, according to the manufacturer’s instructions (Díaz et al., 2015 (link)). Stained cells were analyzed with a FACSAria II flow cytometer (BD Biosciences). The percentage of positive cells and the mean fluorescence intensity (arbitrary units) for a specific marker were calculated using FACSDiva software (BD Biosciences). For each sample, 30.000 events were recorded.
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