Anti cd19 ebio 1d3

BrdU was delivered to mice through drinking water at the concentration of 1mg/ml + 1% glucose. Spleen and lymph nodes were harvested on day 6 post-immunization or infection. Following tetramer enrichment, cells were stained with cell surface antibodies [anti-CD3, anti-CD4, anti-CD19 (eBio (1D3), eBiosciences), anti-CD8α, anti-CD11c (eBiosciences), anti-F4/80 (BM8, Biolegend) and anti-CD5 (53–7.3, BD Pharmingen)] followed by intracellular anti-BrdU (BD Pharmingen) antibody according to the BrdU flow kit protocol (BD Biosciences).

Antibodies used for flow cytometry analysis of surface marker expression on BMDCs are the following: anti-CD11c (N418); anti-CD40 (1C10); anti-CD80 (16-10A1); anti-CD86 (GL1); and anti-I-A/I-E (M5/114.15.2) (all eBioscience). Antibodies used for flow cytometry analysis of cells from mediastinal LNs are the following: anti-CD19 (eBio1D3); anti-F4/80 (BM8) (eBioscience); anti-CD3ε (145-2C11); anti-CD45 (30-F11); anti-CD11c (N418); and anti-I-A/I-E (M5/114.15.2) (BioLegend). For staining of intracellular protein iNOS (C-11) (Santa Cruz Biotechnology), samples were first fixed with IC Fixation Buffer (eBioscience) and stained in Permeabilization Buffer (eBioscience). Samples were acquired on the BD LSR Fortessa and data analyzed using FlowJo software.