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.
ES cells were transiently transfected with flag-tagged mouse NeuroD1 (tagged on N-term). Fixed 24h post transfection.Stained for flag tag (red) to check that some cells express protein. Most protein was in nucleus but some was cytoplasmic. Stained with NeuroD1 N-term antibodies at 1:100. NeuroD1 N-term antibody showed strong and clear staining with similar pattern to the flag staining.(Supplied by Sally Lowell,Edinburgh University)
Fluorescent confocal image of HepG2 cell stained with hNeuroD1-Q30. HepG2 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with hNeuroD1-Q30 primary antibody (1:25, 1 h at 37°C). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:400, 50 min at 37°C). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (7units/ml, 1 h at 37°C). Nuclei were counterstained with DAPI (blue) (10 ug/ml, 10 min). hNeuroD1-Q30 immunoreactivity is localized to vesicles significantly.
Western blot of hNeuroD1-Q30 in HepG2 cell line lysates (35 ug/lane). NEUROD1 (arrow) was detected using the purified antibody.