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Subsequently, the proteins (20 ~ 80 g) were separated by 8%, 10%, or 12% sodium dodecyl sulfated-polyacrylamide gel electrophoresis using voltage mode ranging from 80 to 120 V, and then transferred to 0.22 m of polyvinylidene fluoride membranes (Millipore, Billerica, Mass., USA) using the Mini Trans-Blot electrophoretic transfer cell system (Bio-Rad Laboratories, Inc., Hercules, CA, USA)

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Insert the needle into the vial, flip the vial upside down, and slowly pull the plunger to draw up the prescribed number of units