Anti blimp1 6d3

B cells from C57BL/6 mice were stimulated with LPS (3 μg ml⁻¹) plus IL-4 (4 ng ml⁻¹) in the presence of nil, butyrate (500 μM), propionate (2000 μM), or butyrate (500 μM) plus propionate (2000 μM) for 72 h. Cells were harvested and lysed in Laemmli buffer. Cell extracts containing equal amounts of protein (20 μg) were fractionated through SDS-PAGE (10%). The fractionated proteins were transferred onto polyvinylidene difluoride membranes (Bio-Rad Laboratories) overnight (30 V) at 4 °C. After blocking and overnight incubation at 4 °C with anti-AID (ZA001, Invitrogen), anti-Blimp1 (6D3, eBioscience), anti-acetyl-histone H3 (H3K9ac/K14ac, 17–615, Millipore), anti-histone H3 (601901, BioLegend), or anti-β-Actin mAb (AC-15, Sigma-Aldrich), the membranes were incubated with HRP-conjugated secondary Abs. After washing with PBS-Tween 20, bound HRP-conjugated Abs were detected using Western Lightning^® Plus-Enhanced Chemiluminescence reagents (PerkinElmer Life and Analytical Sciences).