ICC/IF image of ab115135 stained NIH-3T3 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 ab115135 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (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-VCAM1 antibody (ab115135) at 1 µg/mlLane 1 : Spleen (Mouse) Tissue LysateLane 2 : Spleen (Rat) Tissue LysateLane 3 : Thymus (Mouse) Tissue LysateLane 4 : Thymus (Rat) Tissue LysateLane 5 : Lung (Mouse) Whole Cell Lysate - normal tissue (ab29297)Lysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.