Western blot of Rabbit anti-SAE1 antibody shows detection of SAE1. Left lane (-) contains 20 ug human HeLa whole cell protein. Right lane (+) contains 20 ug human HeLa whole cell protein from cells pre-treated with phosphatase inhibitor cocktail to prevent dephosphorylation of the target. Proteins were separated on a 10% SDS-PAGE and transferred onto nitrocellulose. After blocking with 5% milk-TBST 1 hr at room temperature, the membrane was probed with the primary antibody, Anti-SAE1, diluted to 2 g/mL at room temperature for 3 hr followed by washes and reaction with HRP-conjugated secondary and ECL imaging. Personal communication, Xin-Hua Feng, Baylor College of Medicine, Houstin, TX.