Western blot analysis of human and rat HMGB1 using anti-HMGB1, mAb (rec.) (GIBY-1-4) Different amounts of cell extracts from HEK293T cells (3μg, 5μg and 30μg) either transfected with a plasmid coding for rat HMGB1 (lanes 1, 2, 3) or non-transfected (lanes 4, 5, 6), were separated by SDS-PAGE under reducing conditions, transferred to nitrocellulose and incubated with anti-HMGB1,mAb (rec.) (GIBY-1-4) (1μg /ml). Proteins were visualized by a chemiluminescence detection system.