Mouse anti armadillo

Guts were dissected in PBS, fixed for 45 min at room temperature in 100 mM glutamic acid, 25 mM KCl, 20 mM MgSO4, 4 mM sodium phosphate, 1 mM MgCl2, and 4%formaldehyde, washed for 1hr, and incubated with primary antibodies and second antibodies in washing buffer (PBS, 0.5% BSA, 0.1% Triton X-100).
The following primary antibodies were used: rabbit anti-peIF2α antibody (Cell Signaling: 3597, 1:150), rat anti-Delta (gift from Dr. MD Rand, University of Rochester, 1:1000); rabbit anti-pH3 (phosphorylated histone H3, Upstate, 1:1000), mouse anti-β-galactosidase (Developmental Studies Hybridoma Bank, 1:500), rabbit anti-β-galactosidase (Cappel, 1:5000), mouse anti-Armadillo (Developmental Studies Hybridoma Bank, 1: 250)
For Delta antibody staining, guts were fixed using a methanol-heptane method as descried (Lin et al., 2008).
Fluorescent secondary antibodies were purchased from Jackson ImmunoResearch Laboratories. DNA was stained using DAPI. Confocal imaging was performed on a Zeiss LSM700 confocal microscope and processed using ImageJ and Adobe Illustrator.

Testes were dissected in 1× PBS and fixed in 4% formaldehyde for 30 min. For immunostaining, testes were incubated with primary antibodies overnight at 4°C, followed by washes in 1× PBST and incubation with secondary antibodies for two hours at RT. The following primary antibodies were used: rabbit anti-Lid (1:1000; from Julie Secombe, Albert Einstein College of Medicine, Bronx, NY, USA); mouse anti-Armadillo [1:100; developed by Eric Wieschaus, Princeton University, Princeton, NJ, USA, and obtained from Developmental Studies Hybridoma Bank (DSHB)]; rat anti-Vasa (1:100; developed by Allan Spradling and Dianne Williams and obtained from DSHB); rabbit anti-Vasa (1:100; Santa Cruz, sc-30210); chicken anti-GFP (1:1000; Abcam, #13970); rabbit anti-Stat92E (1:800; from Denise Montell, Johns Hopkins School of Medicine, Baltimore, MD, USA); rabbit anti-phospho-Histone H3 (Thr3) (1:200; Millipore, #05-746R); mouse anti-α-spectrin (1:50; obtained from DSHB); mouse anti-γ-tubulin (1:100, Sigma, GTU-88); mouse anti-FasIII (1:50; obtained from DSHB, 7G10); rabbit H3K4me3 (1:200; Cell Signaling, #9751S); rabbit anti-Zfh1 (1:5000; from Ruth Lehmann, Skirball Institute of Biomolecular Medicine, NY, USA). Alexa Fluor 488, 568 and 633-conjugated Goat anti-mouse, anti-rabbit, and anti-rat secondary antibodies were used (1:200; Molecular Probes/Invitrogen).