ab53104 at 1/50 dilution staining NRG1 in human brain tissue by Immunohistochemistry, Paraffin embedded tissue, in the absence and presence of the immunising peptide.
All lanes : Anti-NRG1 antibody (ab53104) at 1/500 dilutionLane 1 : SK N SH (Human neuroblastoma) Whole Cell Lysate Lane 2 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionObserved band size : 100 kDa (why is the actual band size different from the predicted?)Additional bands at : 55 kDa (possible isoform),90 kDa (possible isoform).
ICC/IF image of ab53104 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53104, 5µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.