Multiplex fluorescent western blot for the detection of Vinculin and HSP-90 on HeLa whole cell lysates with Cyanagen IR-BLOT Secondary antibodies
Sample: 2-fold dilution series of HeLa whole cell lysate (abcam®) from 5μg to 0.078 μg of total protein.
Membrane: Trans-Blot® Turbo™ Mini Nitrocellulose Transfer Packs (Bio-Rad). Blocking: 3% ECL™ Blocking Agent (GE Healthcare) in PBS.
Primary antibody: Rabbit anti-Human HSP-90 (Santa Cruz Biotechnology) 1:4000 and Mouse anti-Human Vinculin (Sigma) 1:5000. Secondary antibody:IR-BLOT 700 Goat anti-Rabbit (red) and IR-BLOT 800 Goat anti- Mouse (green)1:25000.
Imager: ODISSEY ® CLX – LI-COR.