Wet Transfer Protocol. Prepare transfer buffer for wet and semi dry transfers based on gel chemistry. Clamp the transfer case closed and submerge it into a transfer tank containing transfer buffer.
Always wear gloves when handling membranes to avoid fingerprints which will negatively affect the results. Pvdf membranes need to be pre wetted in methanol and water before equilibration in transfer buffer. Then soak it in methanol.
In a small flat tupperware like or glass container of 1x transfer buffer wet two blotting sponge pads.
Apparatus used is biorad mini transblot tankwet transfer method. It will transfer the protein as high as 500kda. Ensure you use high molecular weight marker. If using pvdf membrane wet the membrane in methanol for 15 seconds.