Fig. 4
From: Jararaca GPIb-binding protein causes thrombocytopenia during Bothrops jararaca envenomation
(a) Purification profile for UF50 chromatographed on a Source 15PHE column. Adsorbed proteins were eluted with a decreasing linear gradient of (NH4)2SO4 (blue line). The peaks were injected in mice (right panel), and those that induce thrombocytopenia were marked with red and green asterisks. mAu, milli-units of absorbance at 280 nm. (b) Protein purification profile of fraction E analyzed by reversed-phase HPLC. A Venusil XBP C8 column (5 μm, 100 Å, 4.6 × 250 mm, Agela Technologies) was equilibrated with solvent A (0.1% trifluoroacetic acid, TFA, in ultrapure water), and the protein was eluted with a 0–100% increasing linear gradient of solvent B (0.1% TFA/90% acetonitrile) and monitored at 280 nm. (c) Electrophoretic profiles of fraction E in 15% SDS-PAGE, under reducing and non-reducing conditions. Protein E had a single 27.5 kDa chain under non-reducing conditions, and two chains, 15.4 kDa and 12.9 kDa, under reducing conditions. Molecular mass markers are shown in the left side. Gel was silver stained44.
