Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.
Freshly isolated mouse hepatocytes plated on coverslips (2 x105 cells/22-mm glass coverslip) were cultured under normoxic conditions for 6 hr. The cells were then fixed in 2% paraformaldehyde in PBS for 1 hr, and processed for confocal immunofluorescence (red: F-actin, blue: ATP-synthase, green: BNIP3). Fluorescence labeling of BNIP3 accomplished with anti-BNIP3 antibody. Data courtesy of Ruben Zamora, University of Pittsburgh.
Fluorescent confocal image of HepG2 cells stained with BNIP3 (BH3 Domain Specific) antibody. HepG2 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated BNIP3 (BH3 Domain Specific) primary antibody (1:500, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 5 min). BNIP3 immunoreactivity is localized to the cytoplasm of HepG2 cells.
The anti-NIP3 BH3 domain antibody is used in Western blot to detect NIP3 BH3 in Ramos cell lysate (lane 1) and in mouse brain tissue lysate (lane 2).