Rabbit anti dcp 1 asp216

Immunostaining of flat-prepped stage 16 Drosophila embryos was performed using the following primary antibodies: mouse anti-Fas 2 (1:100, clone 1D4, DHSB), mouse anti-axons CNS (1:100, BP 102, DHSB), mouse anti-Futsch (1:1000, 22c10, DHSB), rabbit anti-phospho-JNK (Thr¹⁸³ / Tyr¹⁸⁵) (1:100, Cell Signaling #9251), rabbit anti-Dcp-1 (Asp216) (1:100, Cell Signaling #9578), rabbit anti-GFP tag polyclonal (1:600, Thermo Fisher Scientific), rat anti-Elav (1:1000, clone 7E8A10, DHSB), mouse anti-Repo (1:100, clone 8D12 DHSB), mouse anti-Even-skipped (1:100, clone 3C10, DHSB), mouse anti-Engrailed (1:100, clone 4D9, DHSB) and TRITC-conjugated goat anti-HRP (1:200, Jackson ImmunoResearch #123-025-021).
The secondary antibodies used for detection were: Goat anti-Rabbit IgG (H + L), Alexa Fluor 488 conjugate (A-11008), Goat anti-Rabbit IgG (H + L) Alexa Fluor 555 conjugate (A-21428), Goat anti-Mouse IgG (H + L) Alexa Fluor 488 conjugate (A-11001), Goat anti-Mouse IgG (H + L) Alexa Fluor 555 conjugate (A-21422) and Goat anti-Rat IgG (H + L) Alexa Fluor 555 conjugate (A-21434). All secondary antibodies were used in a dilution of 1:600 and were from Invitrogen.

Experimental procedures for electron and fluorescence microscopy were performed as described earlier (Pertceva et al., 2010 (link)). The primary antibodies were: monoclonal rabbit anti-VASA antibodies from Santa Cruz Biotechnology (1:300), rabbit anti-Dcp-1 (Asp216) from Cell Signaling Technology. The secondary antibodies were goat anti-rabbit conjugated to AlexaFluor-488 (1:500; Invitrogen #A-11001). TRITC-labeled phalloidin (Sigma-Aldrich #P1951) was used at 1:100 dilution to visualize F-actin as described previously (Guild et al., 1997 (link)). The LysoTracker assay was performed as described in Dorogova et al. (2014) (link) (LysoTracker red DND-99 (Invitrogen, Molecular Probes, Basel, Switzerland). Ovaries were embedded with ProLong Gold anti-fade reagent with DAPI. Images were obtained using an AxioImager Z1 microscope with ApoTome attachment (Zeiss), AxioCam MR and AxioVision software (Zeiss, Germany).

Staining procedures were performed as described elsewhere19 (link)20 (link), using mouse anti-Col (1/200; ref. 2 (link); guinea-pig anti-Col (1/5,000; A. Moore, Doshisha University, Kyotanabe, Kyoto, Japan); rabbit anti-H3P (1/200; Upstate Biotechnology); mouse anti-proPO (1/200; T. Trenczel, Justus-Liebig-University Giessen, Giessen, Germany); anti-P1 (1/30; I. Ando, Institute of Genetics, Biological Research Center of the Hungarian Academy of Science, Szeged, Hungary); mouse anti-Antp (1/100), anti-Dlp (1/50), anti-Robo1 (1/10), anti-Robo3 (1/10) and anti-Slit (1/10; Hybridoma Bank); rabbit anti-Robo2 (1/200; B. J. Dickson, Research Institute of Molecular Pathology, Vienna, Austria); and mouse anti-HA (HA11; 1/100; Covance) and rabbit anti-Dcp-1(Asp216) (antibody #9,578, 1/200; Cell Signaling Technology).