All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 4 antibody (ab88239) at 1 µg/mlLane 1 : SW480 (Human colon adenocarcinoma cell line) Whole Cell LysateLane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 5 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)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.
ICC/IF image of ab88239 stained PC12 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 (ab88239, 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) PC12 cells at 5µg/ml.
All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 4 antibody (ab88239) at 1 µg/mlLane 1 : Cerebellum (Mouse) Tissue Lysate Lane 2 : Cerebellum (Rat) Tissue Lysate Lane 3 : Brain (Rat) Tissue LysateLysates/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.