BCHE Antibody / Butyrylcholinesterase (FY13026)

Western blot analysis of BCHE using anti-BCHE antibody. Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-BCHE antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. A doublet is observed at ~85-90 kDa, higher than the predicted 68 kDa molecular weight. This shift is well documented and results from extensive N-linked glycosylation of the mature secreted enzyme, which adds ~15-20 kDa of carbohydrate mass. Variability in glycan composition or processing produces the characteristic doublet pattern seen for BCHE in multiple mammalian cell types.