All lanes : Anti-VGF antibody (ab74140) at 1/500 dilutionLane 1 : Extracts from RAW264.7 cellsLane 2 : Extracts from RAW264.7 cells with immunising peptide at 5 µgLysates/proteins at 5 µg per lane.
Immunohistochemical analysis of VGF in paraffin embedded human lung carcinoma tissue using ab74140, at a 1/50 dilution, in the presence (right panel) or absence (left panel) of immunising peptide.
ICC/IF image of ab74140 stained PC12 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 (ab74140, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (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.