Anti-EEA1 antibody [1G11] - Early Endosome Marker (ab70521)

ICC/IF image of ab70521 stained HepG2 cells. The cells were fixed with 4% PFA (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 (ab70521 at 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 Goat anti-Mouse 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). This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml.

ICC/IF image of ab70521 stained HepG2 cells. The cells were 4% PFA 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 (ab70521, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight^® 488 oat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.

ab70521 staining EEA1 in human THP-1 cells by Immunocytochemistry/ immunofluorescence. Cells were formaldehyde fixed, permeabilized in TritonX100 and blocked in 1% BSA for 1 hour at 25°C. The primary antibody was diluted 1/200 (PBS + Tween20 + BSA 1%) and incubated with sample for 1 hour at 25°C. A 1.4 nm Gold (ultra-small) and Alexa Fluor^® 594 conjugated goat polyclonal to mouse IgG1, diluted 1/500 was used as secondary. The red staining in image show EEA1 staining whilst green cytosolic staining corresponds to StarD5.

See Abreview