ab654 at 1/500 dilution staining GLUT4 in mouse muscle by immunohistochemistry (frozen sections). Sections were methanol fixed prior to blocking in 5% goat serum for 1 hour at 25°C and then incubated with ab654 for 1 hour at 25°C. A goat polyclonal to rabbit Ig, diluted 1/500, was used as the secondary antibody.See Abreview

ICC/IF image of ab654 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 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 (ab654, 1/1000 dilution) 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.