Western blot using Rabbit-anti-SAE1 pS185 antibody shows detection of phosphorylated SAE1. Left lane (-) contains 20ug human HeLa whole cell protein. Right lane (+) contains 20ug 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 diluted to 1:1,000 at room temperature for 3 hr followed by washes and reaction with HRP-conjugated secondary and ECL imaging.
