Twenty-five microframs of lysate (in β-mercaptoethanol-containing buffer) was resolved in a 9% (or 4 20% gradient for APOB) polyacrylamide gel and analyzed by Western blot.
Brain, liver, and spleen homogenates (10%) prepared in lysis buffer were resolved by SDS-PGE and subjected to western blotting using specific antibodies as described in previous reports [30].
Cell lysates extracted in RIPA buffer were resolved by 3 8% SDS-PAGE.
Total cell protein, extracted using RIPA buffer, were resolved by SDS-PAGE (8%) in parallel duplicated gels, transferred to nitrocellulose membranes and incubated with primary antibody ((Phospho-EGFR Tyr-1045, or total EGFR; Cell Signalling Technology) diluted 1/1000 in 5% BSA in TBS-T overnight at 4°C.
Proteins in Laemmli sample buffer were resolved by SDS-PAGE (10%) and transferred to PVDF Immobilon-P membrane.
Cells lysates, prepared by dissolving washed cells directly in SDS sample buffer, were resolved by SDS-PAGE with indicated percentage of acrylamide.
Whole-cell protein extracts prepared in Laemmli sample buffer were resolved by SDS-PAGE using NuPage 4 12 % gels (Invitrogen), transferred to Invitrolon polyvinylidene difluoride membranes (Invitrogen), and probed with primary and horseradish peroxidase-conjugated secondary antibodies.
Whole-cell protein extracts prepared in Laemmli sample buffer were resolved by SDS-PAGE using 4% to 12% polyacrylamide gels (NuPage; Invitrogen Corp ., transferred to PVDF membranes (Invitrolon; Invitrogen Corp ., and probed with primary and horseradish peroxidase-conjugated secondary antibodies.
Protein samples boiled in SDS-sample buffer were resolved on a 6% SDS-PAGE, transferred onto a nitrocellulose membrane, and then blotted with a polyclonal anti-c-Met antibody (Santa Cruz Biotechnology, Santa Cruz, CA), monoclonal anti-phospho-c-Met antibody (Cell Signaling Technology, Danvers, MA), or monoclonal anti-tubulin antibody (Thermo Scientific, Fremont, CA).
