Figure 1

Production and characterization of murine adiponectin oligomers. A) Western blot of gel fractionated adiponectin oligomers from a full-length preparation. The highest molecular weight form elute first (left to right). Fractions representing HMW, LMW, and trimer adiponectin were pooled for their respective purifications. Dotted lines mark typical fraction cut-offs for pooling. The band for 28 kDa molecular weight marker (from the Bio-Rad 2 color ladder) is noted. The less intense slower band labeled is N-glycosylated – both bands were included in intensity quantification. B) Quantitation of oligomer distribution in purified preparations used in experiments; data are presented as 0^th order, 2-neighbor smoothed curves for clarity. Percent distributions were calculated from raw intensity data with peaks separated by fraction intensity peak valleys of the full-length form. C) Dynamic light scattering measurements of purified adiponectin oligomers yield estimated Stokes radii and molecular weights. Murine IgGs, albumin, and linear known-length dextran molecules were measured for comparison. All of the proteins – including adiponectin – followed a globular protein relationship while the dextrans fit a linear molecule relationship.