Mouse tubulin primary antibodies

To perform western blots, fly heads were homogenized in SDS sample buffer with a Pellet Pestle (Kimble/Kontes). The proteins were fractionated by SDS-PAGE and transferred to Immobilon-P transfer membranes (Millipore) in Tris-glycine buffer. The blots were probed with mouse Tubulin primary antibodies (1:2000 dilution, Developmental Studies Hybridoma Bank), mouse Rh1 antibodies (1:2000 dilution, Developmental Studies Hybridoma Bank) and Rat anti-INAD (1:2000, C. Montell lab), followed by IRDye 680 goat anti-mouse IgG (LI-COR) and IRDye 800 goat anti-Rat IgG (LI-COR) as the secondary antibodies. The signals were detected with the Odyssey infrared imaging system (LI-COR).

Western blotting was performed as described [39 (link)]. Briefly, 20 fly eyes were dissected and homogenized in SDS sample buffer with a Pellet Pestle (Kimble/Kontes). The proteins were fractionated by SDS-PAGE and transferred to Immobilon-P transfer membranes (Millipore) in Tris-glycine buffer. The blots were probed with Mouse Tubulin primary antibodies (1:2000 dilution, Developmental Studies Hybridoma Bank), Rabbit ERK (1:1000, Cell Signaling Technology), Mouse β-Actin primary antibodies (1:1000 dilution, Santa Cruz Biotechnology) and Rabbit RACK1 antibodies (1:1000 dilution, Dr. J. Kadrmas lab) followed by IRDye 680 goat anti-Rabbit IgG (LI-COR) and IRDye 800 goat anti-Mouse IgG (LI-COR) as the secondary antibodies. The signals were detected with the Odyssey infrared imaging system (LI-COR).