Immunofluorescence Labeling of Drosophila Tissues

The chitin-binding probe was prepared in our laboratory as described [91 (link)]. Chitin-546 was used at 1:10 dilution. The following primary antibodies were used: Chicken anti-GFP (1:200) (Abcam), Guinea Pig anti-Uif (1:300) [69 (link)], Mouse anti-Gasp (1:5) (DSHB), Guinea Pig anti-Cor (1:500) [66 (link)], Rabbit anti-Verm (1:100) [27 (link)], Rabbit anti-Kune-kune (1:200) [43 (link)], Mouse anti-DLG (1:1) (DSHB), Mouse anti-Spectrin (1:10) (DSHB), Mouse anti-Cnx99A (1:5) (DSHB), Rabbit anti-GM130 (1:200) (Abcam), Mouse anti-KDEL (1:200) (Abcam), Rabbit anti-Rab5 (1:200) (Abcam), Rat anti-Rab11 (1:100) [92 (link)], and Mouse anti-Rab7 (1:10) (DSHB). Antibodies generated in this study: Rabbit anti-Osi19 (1:300), Rat anti-Osi15 (1:25), Guinea Pig anti-Osi9 (1:10).
The following secondary antibodies (Invitrogen life technologies) were used: AlexaFluor 488 anti-chicken IgG, anti-rat IgG, anti-mouse IgG, anti-rabbit IgG, anti-guinea pig IgG; AlexaFluor 543 anti-rat IgG, anti-rabbit IgG, anti-guinea pig IgG, anti-mouse IgG; AlexaFluor 594/546 anti-mouse IgM; AlexaFluor 647 anti-rat IgG, anti-mouse IgG, anti-rabbit IgG, anti-guinea pig IgG; AlexaFluor 647 plus anti-mouse, anti-rabbit; AlexaFluor 488 plus anti-mouse, anti-rabbit), and AlexaFluor 405 anti-Guinea Pig. All secondary antibodies were used at 1:200 dilution.