• Open apparatus by pushing toggles (A) symmetrically towards the centre so that the cathode lifts off to reveal the anode
• Load three sheets of soaked filter paper, one at a time, onto the plate. Use a glass rod as a mini rolling pin to squeeze out any bubbles between sheets. Add soaked PVDF membrane after snipping top rh corner ( this will mark the side to which protein has transferred. Add gel. Then four further sheets of soaked filter paper, one at a time, squeezing out bubbles at each stage.
• Up to six stacks can be loaded on each machine
• Set current to 100mA per stack ( 600mA for a full load of 6- an initial voltage of 10V is usually sufficient). During the run, increase the voltage to restore the initial voltage, but never exceed 25V.
• When retrieve PVDF membrane at the end, show students the transient outline of bands as the membrane dries ( membrane is hydrophobic so dries more quickly than protein).
• After ECL, wrap in gel film with untidy ends behind (where "in front" has the cut  corner upper right). Place membranes in film cassette first, orientated so that the cut corners are upper right. Then put film on top.

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