Formalin-fixed and paraffin-embedded human Spleen tissue reacted with NANOG Antibody , which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Fluorescent confocal image of HeLa cell stained with NANOG Antibody. HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with NANOG 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). NANOG immunoreactivity is localized to Nucleus significantly.
Fluorescent confocal image of SY5Y cells stained NANOG antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated NANOG 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). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (5.25 mu M, 25 min). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 3 min). Nanog immunoreactivity is localized mainly to the nuclei of the SY5Y cells.
Western blot of anti-NANOG Antibody in K562 cell line lysates (35 ug/lane). NANOG (arrow) was detected using the purified antibody.Western blot of NANOG (arrow) using rabbit polyclonal NANOG Antibody. 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the NANOG gene (Lane 2) (Origene Technologies).
NANOG Antibody flow cytometry of HepG2 cells (right histogram) compared to a negative control cell (left histogram). FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.