All lanes : Anti-Calnexin antibody - ER Membrane Marker (ab140818) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Brain (Mouse) Tissue LysateLane 3 : Brain (Rat) Tissue LysateLane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 5 : Brain (Mouse) Tissue LysateLane 6 : Brain (Rat) Tissue LysateLysates/proteins at 10 µg per lane.SecondaryLanes 1 - 3 : Anti-Chicken IgY VHH Single Domain Antibody (HRP) (ab191865) at 1/50000 dilutionLanes 4 - 6 : Goat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/20000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab140818 stained HeLa cells. The cells were 4% formaldehyde 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 ab140818 at 5 overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- chicken IgY (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.This antibody also gave a positive result in the following 4% formaldehyde fixed (10 min), and 100% methanol fixed (5 min) cell lines at 5µg/ml, HepG2, MCF-7, HeLa.
All lanes : Anti-Calnexin antibody - ER Membrane Marker (ab140818) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell LysateLane 4 : Brain (Mouse) Tissue LysateLane 5 : Brain (Rat) Tissue LysateLane 6 : Liver (Mouse) Tissue LysateLane 7 : Liver (Rat) Tissue LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.