Mouse anti tfr

Lysates from whole-cell extracts, cell-surface proteins, or intracellular proteins were subjected to gel electrophoresis. Proteins were then transferred to PVDF membranes (Bio-Rad, Burlington, ON). The membranes were blocked in a solution of TBS-T containing 4% BSA or 3% milk for 30 minutes at room temperature and then incubated at room temperature with the primary antibody for 1h. Rabbit anti-CD71 (1:1000), rabbit anti-Glut1 (1:1000), and rabbit anti-EGFR (1:1000) were purchased from GeneTex (Irvine, CA), goat anti-EGFR, mouse anti-TfR
(1:400, 1:1000), and mouse anti-CHC (1:1000) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-β1 integrin (1:1000), rabbit anti-pAMPK (1:500), rabbit anti-pACC (1:500), rabbit anti-pan-actin (1:1000), and rabbit anti-total ERK (1:1000) were purchased from Cell Signaling Technology (Danvers, MA). Mouse anti-β1-integrin (1:400) was purchased from Calbiochem (Billerica, MA). Secondary antibodies were goat anti-rabbit, goat anti-mouse, (Jackson Immunoresearch, West Grove, PA). Streptavidin-POD (1:1000) was purchased from Roche.