Alexa505

Larval tissues were fixed for 20 min in 4% formaldehyde/phosphate-buffered saline (PBS) and immunostained as described (Bello et al. 2003 (link)). The primary antibodies used were rabbit anti-aPKC (1:500; Santa Cruz Biotechnology), mouse anti-Mira (1:50; gift of A. Gould), rat anti-pH3 (1:500; Abcam), chick anti-GFP (1:2000; Abcam), rabbit anti-Dpn (1:100; gift of Y.N. Jan), guinea pig anti-Dpn (1:1000; gift of James Skeath), rat anti-Pros (1:50; gift of F. Matsuzaki), rabbit anti-Ase (1:50; gift of F. Matsuzaki), rat anti-CycE (1:500; gift of Helena Richardson), mouse anti-CycA (1:50; Developmental Studies Hybridoma Bank), mouse or rat anti-Elav (1:100; Developmental Studies Hybridoma Bank), rabbit anti-Insc (1:1000; gift of W. Chia), rabbit anti-Myc (1:100; Santa Cruz Biotechnology), guinea pig anti-Nerfin-1 (1:5000; gift of A. Kuzin), rabbit anti-RFP (1:100; Abcam), and mouse anti-Fib (1:200; Abcam). Secondary goat antibodies conjugated to Alexa488, Alexa568, Alexa650, and Alexa505 (Molecular Probes) were used at 1:200. Images were collected on a Leica SP5 confocal microscope, and all images shown are single sections unless otherwise stated.