ICC/IF image of ab134015 stained SKNSH 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 (ab134015, 0.5µg/ml) overnight at +4°C. The secondary antibody (green) was ab46947, DyLight® 488 goat anti-chicken IgY (H+L) used at a 1/250 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.
Immunohistochemical analysis of dissociated cell cultures of neonatal mouse brains, labelling NSE-2 (green staining) in neurons using ab134015 at 1/2000 dilution. Cultures were counter-stained with a rabbit anti-GFAP to localize astrocytes, as well as with DAPI (blue staining) to localize nuclei.
Anti-NSE antibody (ab134015) at 1/5000 dilution + Adult mouse brain solubilized with Lamelli cracking buffer