Drosophila embryos were dechorionated in 2.15% sodium hypochlorite and then either heat-fixed in a salt solution (Miller et al., 1989 (link); Figs. 9 and S4 ) or fixed for 20 min in 4% formaldehyde in a 1:1 PBS:heptane mixture (Fig. S2 ). Primary antibodies used under heat fixation were anti-HA (rat monoclonal, 3F10; 1:500; Roche), anti-Cno (rabbit 1:1,000, a gift from Mark Peifer, University of North Carolina, Chapel Hill, NC, USA), anti-DE-cadherin (rat monoclonal DCAD2 1:25; Developmental Studies Hybridoma Bank). Under heptane fixation: anti-HA (mouse 16B12 1:100; Abcam). For immunostaining in Fig. S1 , ovaries were fixed in 5% formaldehyde in phosphate buffer for 10–12 min. Primary antibodies used for immunostaining were: anti-HA (rat monoclonal 3F10, 1:600; Roche); anti-Arm (mouse monoclonal N2 7A1, 1:100; DSHB); anti-α-spectrin (rabbit polyclonal, 1:1,000, gift from Daniel Branton, Harvard University, Cambridge, MA, USA); anti-GFP (mouse monoclonal JL-8, 1:400; Clontech). Secondary antibodies were used at a concentration of 1:400 (Invitrogen).
Anti de cadherin
Manufactured by Developmental Studies Hybridoma Bank
Sourced in United States
Anti-DE-cadherin is a monoclonal antibody that binds to the DE-cadherin protein. DE-cadherin is a cell-cell adhesion molecule found in Drosophila. The antibody can be used to detect and study the localization and expression of DE-cadherin in various biological samples.
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2 protocols using anti de cadherin
1
Immunostaining of Drosophila Embryos and Ovaries
2
Immunofluorescence Staining of Ovaries and Testes
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Ovaries and testes were fixed in 5% formaldehyde (Polysciences, Inc.) for 10 min and stained as described in König and Shcherbata (2013) (link). We used the following mouse monoclonal antibodies: anti-adducin (1:50), anti-lamin C (1:50), anti-arm (1:50), anti-FasIII (1:50), rat monoclonal antibody anti-DE-cadherin (Developmental Studies Hybridoma Bank), guinea pig anti-Tj (1:3000, D. Godt), rabbit anti-vasa (1:5000, gift from R. Pflanz), anti-β-Gal (1:3000, Cappel), and anti-GFP-directly conjugated with AF488 (1:3000, Invitrogen), Alexa 488, 568, or 633 goat anti-mouse, anti-rabbit (1:500, Molecular Probes), goat anti-rat Cy5 (1:250, Jackson Immunoresearch). Images were obtained with a confocal laser-scanning microscope (Zeiss LSM700) and processed with Adobe Photoshop.
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