All lanes : Anti-HUNK antibody (ab46726) at 1 µg/mlLane 1 : MDA MB 361 (Human breast adenocarcinoma cell line) Whole Cell LysateLane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell LysateLane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 4 : ZR-75-1 (Human breast carcinoma cell line) Nuclear lysate (ab14916)Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
ICC/IF image of ab46726 stained human MCF7 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab46726, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
ab46726 staining HUNK in mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/1000 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo488 conjugated goat polyclonal to rabbit IgG was used undiluted as secondary. The picture shows the staining obtained in the mouse cortex and located in the soma and processes of some neurons.See Abreview