
ICC/IF image of ab87353 stained RAW246.7 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 (ab87353, 10µg/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.

All lanes : Anti-15 Lipoxygenase 1 antibody (ab87353) at 1 µg/mlLane 1 : Kidney (Mouse) Tissue LysateLane 2 : Heart (Mouse) Tissue Lysate Lane 3 : Small Intestine (Mouse) Tissue Lysate Lane 4 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell LysateLane 5 : Brain (Mouse) Tissue LysateLane 6 : Forebrain (Mouse) Tissue Lysate Lane 7 : EL4 (Mouse lymphoma cell line) Whole Cell Lysate (ab7183)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.