Fitc conjugated anti cd19 antibody

A BD FACSCelesta^TM flow cytometer was used to detect cells. The antibodies used in the experiments included AF488-conjugated anti-CD4 antibody (BD Biosciences), FITC-conjugated anti-CD19 antibody (BD Biosciences), BV510-conjugated anti-CXCR3 antibody (BD Biosciences) and phycoerythrin-conjugated anti-TIGIT antibody (BD Biosciences), PE-CF594-conjugated anti-CD8 antibody (BD Biosciences). These were surface antibodies, and were detected after staining in the dark for 30 min. Phycoerythrin-conjugated anti-forkhead box P3 (anti-Foxp3) antibody (BD Biosciences), BV421-conjugated anti-interleukin 4 (anti-IL-4) antibody (BD Biosciences), PerCP-Cy™5.5 -conjugated RoRrt antibody (BD Biosciences), APC-Cy7-conjugated anti-IFN-γ antibody (BD Biosciences) and AF647-conjugated anti-GL-7 antibody (BD Biosciences) are intracellular antibodies, and the cell membrane needed to be disrupted for 40 min prior to staining.

The signaling pathways involved in the development and functions of B cells were assessed using flow cytometry with the same protocol used to assess the DNA damage (described above). The only difference was the antibody used along with an FITC-conjugated, anti-CD19 antibody (BD Biosciences). For the assessment, 25 × 10³ cells were stained using the following Alexa Fluor^® 647 conjugated antibodies from BD Biosciences: phospho-NF-κB (Ser529; clone B33B4WP; Thermo Fisher Scientific), phospho-ERK1/2 (clone 20A), phospho-p38 (clone pT180/py182), and phospho-STAT5 (clone pY694). Data were acquired with the BD Accuri™ C6 flow cytometer. The gating strategy was as follows: cells were identified as CD19+ cells, and the cells positive for the aforementioned targets were identified in this CD19+ population. The data were analyzed using FCS Express™ 6 software (De Novo Software).