WB using Anti-Hif3A antibody shows detection of a band ~72 kDa corresponding to mouse Hif3A (arrowhead). Approximately 10 ug of a CoCl treated 3T3 cell lysate (lane 1) and control 3T3 cell lysate (lane 2) were separated by 4-20% SDS-PAGE and transferred onto nitrocellulose. Treatment of exponentially growing 3T3 cells with 130 µM CoCl for 6 h at 37°C effectively mimics hypoxia. Primary antibody was diluted to 1:1, 600. IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX was used at 1:10000.