Immunoprecipitations using perilipin and vimentin antibodies and briefly AIM-stimulated preadipocytes.

<p>Mabs Peri112.17, Vim3B4 and control VE-cadherin were used. Immunoprecipitations (IPs) were analyzed by SDS-PAGE and CB staining (<b>a</b>) and corresponding WB using pab Peri-hNT (<b>b</b>). Lane 1: control unspecific binding of cell lysate to beads, i.e. magnetic beads after incubation with lysate and PBS washing; lane 2: bound lysate material of perilipin antibody beads; lane 3: control unbound supernatant of perilipin antibody beads; lane 4: bound lysate material of vimentin antibody beads; lane 5: control unbound supernatant of vimentin antibody beads; lane 6: bound lysate material of VE antibody beads; lane 7: control unbound supernatant of VE antibody beads; lane 8: control magnetic beads after incubation with crosslinked lysate; lane 9: bound crosslinked lysate material of perilipin antibody beads; lane 10: bound crosslinked lysate material of vimentin antibody beads; lane 11: bound crosslinked lysate material of VE antibody beads. Positions of molecular weight marker proteins are given on the left margin. Of importance, perilipin and notably vimentin antibodies precipitate and co-precipitate perilipin (<b>lanes 2,4;</b> perilipin position at 65 kD is marked at left margin by an arrow) in contrast to the negative control antibody (<b>lane 6</b>). In addition, similar positive reactions are obtained with crosslinked lysate by detecting crosslinked perilipin (<b>lanes 9,10</b>) whereas the control antibody shows no reaction with crosslinked material at all (<b>lane 11</b>).</p>