Figure 4

Validation of anti-G72 antibodies using western blotting and LI-COR Odyssey ^® Infrared Imaging System. (A) Western blotting was performed using rabbit polyclonal antibodies anti-G72 #1410 (left) or anti-G72 #1411 (right). Lane 1: total protein lysate from mock-transfected HEK-293 cells. Lane 2: total protein lysate from His-G72-transfected HEK-293 cells. Lane 3: Eluate from immunoprecipitation (IP) of mock lysate with anti-G72 #1411. Lane 4: Eluate from IP of His-G72 lysate with anti-G72 #1411. A strong signal for His-G72 was detected at ~22 kDa (lanes 2 and 4), with weaker bands around 18 kDa and below. Additional bands in lanes 3 and 4 (weak signal at 25 kDa and strong signal at 50 kDa) represent the light and heavy chain of the antibody used for IP. (B) G72-specific antibody #1410 (upper panel, green) and a commercial anti-His antibody (middle panel, red) detect the same protein bands on Western blots (merged image, lower panel, yellow) in total protein lysates from HEK-293 cells overexpressing His-G72. Lane 1 & lane 2: as described in A.