Anti cd16 bv650

Flow cytometry was employed to examine the immunophenotype of lymphocytes found in peripheral blood. The process involved obtaining a whole blood sample, which was then treated with a set of monoclonal human antibodies comprising anti-CD4 BV421, anti-CD3 PerCp, anti-CD8 BV605, anti-CD19 FITC, anti-CD45 Alexa Fluor 700, anti-CD56 BV650, anti-CD16 BV650, anti-TLR-2 APC, anti-TLR-4 PE, anti-TLR-7 PE, anti-TLR-8 APC, anti-TLR-3 PE, and anti-TLR-9 APC (Biolegend, San Diego, CA, United States). Following the antibody staining, a lysing buffer (BD, Franklin Lakes, NJ, United States) was applied to eliminate red blood cells, and the resulting cells were thoroughly washed and evaluated using a CytoFLEX LX instrument (Beckman Coulter, Indianapolis, IN, United States). Subsequently, data analysis was carried out utilizing the Kaluza Analysis program (sample analysis in Figure 1—CLL and Figure 2—CVID) The CytoFLEX LX flow cytometer was internally quality controlled using CytoFLEX Ready to Use Daily QC Fluorosphere reagents (Beckman Coulter, Indianapolis, IN, United States).

Flow cytometry evaluated the percentage of TLR-9-positive CD4+/CD8+/CD19+ lymphocytes in a peripheral blood sample. The whole blood sample was incubated with human monoclonal antibodies. These antibodies included anti-CD45 AF700, anti-CD4 BV421, anti-CD8 BV605, anti-CD19 FITC, anti-CD56 BV650, and anti-CD16 BV650, as well as anti-CD3 PerCp and anti-TLR-9 APC (BioLegend, San Diego, CA, USA). After the incubation stage, the samples were treated with a previously prepared lysing solution to remove red blood cells, and then the samples were washed twice. The thus prepared samples were evaluated with the CytoFLEX LX instrument (Beckman Coulter, Indianapolis, IN, USA). The obtained data were analyzed using the Kaluza Analysis program. An exemplary analysis can be found in Figure 1.