For 1D-SDS-PAGE, Protein were fractionated on SDS-PAGE mini gel slab gel (size, AE-6530 mPAGE,ATTO CORPORATION). SDS-PAGE were performed on 12.5 polyacrylamide gel. Whole protein were measured by BCA protein assay. An equal amount of whole protein 50 mg. were loaded. To estimate size of proteins, broad range molecular weight protein standard marker was used (V8491, Promega). The upper and lower reservoirs of the electrophoresis apparatus were filled with electrophoresis buffer
(0.025M Tris-Cl, 0.192M. Glycine, 0.1 SDS). The individual proteins are separated eletrophoretically at constant current of 15 mA of constant current for stacking gel until bromophenolblue tracking dye enters the sepatating gel, and then the current was increased to 20 mA until the tracking dye has reached the end of separating gel.