Goat anti mouse f ab fragment

Primary antibodies used for immunohistochemistry were: rat anti-N-Cadherin extracellular domain (DN-Ex #8; 1:20^{88 (link)}, 1997, DSHB); rabbit anti-GFP (1:1000; Molecular Probes); and mouse anti-CD2 (1:1000; Molecular Probes). Secondary antibodies used were as follows (all 1:300): goat anti-rabbit F(ab)′ fragment coupled to Alexa 488 (Molecular Probes), goat anti-mouse F(ab)′ fragment coupled to Alexa 568 (Molecular Probes), goat anti-mouse F(ab)′ fragment coupled to Alexa 568 highly cross-absorbed (Molecular Probes), goat anti-rat F(ab)′ fragment coupled to Alexa 568 (Molecular Probes), goat anti-rat F(ab)′ fragment coupled to Alexa 647 (Molecular Probes) and Toto-3 (1:2000, Molecular Probes). Immunostaining of brains of adult flies and pupae were carried out essentially as described previously^{89 (link)} with the following exceptions: (1) adult brains were fixed in 2% PFA for 90 min, and (2) for the dissection of the pupal brains, the pupal cases were open, 2% PFA was added, and the brains were allowed to fix for 10 min before further dissection in 2% PFA. The overall time of fixation in 2% PFA was 90 min. Fluorescent samples were analyzed using a Zeiss Meta510 confocal microscope.