All lanes : Anti-ESAM antibody (ab74777) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431) Lane 2 : Human liver tissue lysate - total protein (ab29889) Lane 3 : Human kidney tissue lysate - total protein (ab30203) Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 6 : Lung (Human) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Annexin A2 overexpression 293T lysate (whole cell) (ab94080) at 1/5000 dilution
The ESAM protein has a predicted molecular weight of 41 kDa. The first 29 amino acids of the ESAM sequence act as a signal sequence. The band at 37 kDa may represent the cleaved form of the ESAM protein.
ICC/IF image of ab74777 stained MCF7 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 (ab74777, 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) HepG2 cells at 5µg/ml, and in 100% methanol fixed (5 min) HepG2 and MCF7 cells at 5µg/ml.