Purification of 8972-Da protein from murine plasma. Results from GVHD (A,C,E) and control (B,D,F) plasma are shown. (A,B) SELDI-TOF analyses of HPLC fraction 31.0 to 31.5 minutes derived from GVHD (A) and control (B) samples are shown. The box identifies a peak of 8972 Da that is overexpressed in GVHD (A), but not detectable in control plasma (B). (C,D) 2DE gel images of GVHD (C) and control (D) samples are shown. Plasma proteins (70 μg) of HPLC-purified fractions (pool of 31.0- to 31.5-minute fraction) from GVHD and control samples, respectively, were separated on pH 3-11 IPG in the first dimension, followed by SDS-PAGE on 8% to 20% gradient gel in the second dimension. The separated proteins were stained with silver nitrate. Insert is magnified in panels E,F. (E,F) Detailed views with apparent molecular mass of between 6500 and 14 400 Da in the 2DE gel images of GVHD (C) and control (D) samples are shown. Arrowhead indicates the candidate spot for the 8972-Da peak in GVHD (E) sample, which is not apparent in the control (F) sample.