Tritc conjugated goat anti rabbit secondary antibody

Serial sections (4 µm thick) were cut from 10% formalin-fixed, paraffin-embedded tumor tissue blocks followed by deparaffinization, rehydration and heat-induced epitope retrieval. Sections were simultaneously incubated with primary antibodies against Hep Par-1 and CK19. Then, sections were stained for 1 h at 37 ℃ in the dark with TRITC-conjugated goat anti-rabbit secondary antibody (111-026-045; Jackson ImmunoResearch, USA) and FITC-conjugated goat anti-mouse secondary antibody (115-096-003; Jackson ImmunoResearch). Sections were counterstained for 2 min with 4',6-diamindino-2-phenylindole (C0065; Solarbio, China) to label nuclei, and analyzed under a fluorescence microscope (BX51, Olympus, Japan).

WT and Bok-deficient neurons grown on 13-mm coverslips were fixed with 4% paraformaldehyde for 15 min, permeabilized in PBS containing 0.1% Triton X-100, washed three times with PBS, and blocked for 1 h in 5% goat serum in PBS. Neurons were then incubated for 2 h with an anti-KDEL antibody diluted 1:100 (10C3, SPA827, Assay Designs) in 5% serum in PBS. Primary antibodies were detected using a 1:250 dilution of TRITC-conjugated goat anti-rabbit secondary antibody (Jackson ImmunoResearch, Plymouth, PA, United States) for 1 h. Coverslips were then transferred to glass slides with DAPI (4′,6-diamidino-2-phenylindole) mounting medium and sealed around the edges with clear varnish. Images of stained cells for quantification were captured with the LSM 7.10 confocal microscope equipped with a 63 × 1.4 NA oil immersion objective (Carl Zeiss). All microscope settings including laser intensity and scan time were kept constant for the whole set of experiments. All images were processed and analyzed using MetaMorph Software version 7.5 (Universal Imaging Co.), and the data presented as fluorescence intensity in arbitrary units (AU).