ICC/IF image of ab75491 stained Malme-3M 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 ab75491 at 5µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488 (ab96899) 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.
All lanes : Anti-NFKBIL1 antibody (ab75491) at 1 µg/mlLane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate Lane 4 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate Lane 5 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell LysateLane 6 : THP1 (Human acute monocytic leukemia cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.