Western blot - Anti-Interferon beta antibody (ab85803)
All lanes : Anti-Interferon beta antibody (ab85803) at 1 µg/ml
Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 2 :WI-38 whole cell lysate (ab3960) Lane 3 : Human liver tissue lysate - total protein (ab29889) Lane 4 : Lung (Human) Tissue Lysate Lane 5 : Human kidney tissue lysate - total protein (ab30203) Lane 6 : Human spleen tissue lysate - total protein (ab29699)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
The Interferon Beta protein sequence contains both a phosphorylation and a glycosylation site (Swissprot). Post-translation modifications could account for the migration of the band at a higher molecular weight than predicted.
ICC/IF image of ab85803 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 (ab85803, 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.