Anti-HE4 antibody (ab85179)

All lanes : Anti-HE4 antibody (ab85179) at 1 µg/ml

Lane 1 : Lung (Human) Tissue Lysate
Lane 2 : Human ovary tissue lysate - total protein (ab30222)

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 13 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?


Exposure time: 16 minutes


Human WAP four-disulfide core domain protein 2 precursor (HE4) contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.

ICC/IF image of ab85179 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 (ab85179, 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. This antibody also gave a positive result in 4% PFA fixed (10 min) MCF7 cells at 1µg/ml.